Abstract:
An efficient architecture for an interpolator ( 100 ) disposed to process oversampled data is disclosed herein. The interpolator ( 100 ) includes an input divider circuit ( 104 ) configured to receive an input data word over an input data line. A register ( 108 ) is provided for latching the divided input data word from the divider ( 104 ). The divided input data word is added within a summer ( 112 ) to a latched divided data word from the register, thereby forming a summed data word. A multiplexer ( 116 ) produces an interpolated output by multiplexing the summed data word with an input data word. In a preferred implementation, the register ( 108 ) is latched at a first clock rate, and the multiplexer ( 116 ) is clocked at twice the first clock rate. The efficient filter architecture allows interpolation to be performed in the absence of multipliers, and in a manner using filter coefficients equivalent to powers of two. This enables the interpolator ( 100 ) to be realized inexpensively, and renders the filter particularly suitable for implementation within integrated circuits.

Description:
FIELD OF THE INVENTION  
         [0001]    The present invention relates generally to mammalian α-N-acetylglucosaminidase and to genetic sequences encoding same and to the use of these in the investigation, diagnosis and treatment of subjects suspected of or suffering from α-N-acetylglucosaminidase deficiency.  
           [0002]    Bibliographic details of the publications referred to by author in this specification are collected at the end of the description. Sequence Identity Numbers (SEQ ID NOs.) for the nucleotide and amino acid sequences referred to in the specification are defined following the bibliography.  
           [0003]    Throughout this specification, unless the context requires otherwise, the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.  
         BACKGROUND TO THE INVENTION  
         [0004]    The increasing sophistication of recombinant DNA technology is greatly facilitating the efficacy of many commercially important industries including areas of medical and pharmaceutical research and development. The ability to purify native proteins and subsequently clone genetic sequences encoding these proteins is an important first step in the development of a range of therapeutic and diagnostic procedures. However, practitioners have faced many difficulties in purifying target molecules to an extent sufficient to determine amino acid sequences to permit the development of oligonucleotide probes to assist in the cloning of genetic sequences encoding the target molecules.  
           [0005]    Such difficulties have been particularly faced in the research and development of lysosomal enzymes. An important lysosomal enzyme is α-N-acetylglucosaminidase (EC 2.1.50). This enzyme acts as a exoglycosidase in lysosomes to hydrolyse the terminal α-N-acetylglucosamine residues present at the non-reducing terminus of fragments of heparan sulphate and heparin (Hopwood, 1989). A deficiency in this lysosomal hydrolase is responsible for the pathogenesis of Sanfilippo B (Mucopolysaccharidosis type IIIB [MPS-IIIB]) syndrome (von-Figura and Kresse, 1972; O&#39;Brien, 1972). This is an autosomal recessive disorder of glycosaminoglycan catabolism leading to storage and excretion of excessive amounts of heparan sulphate and a variety of clinical phenotypes, but classically presenting with progressive mental retardation in conjunction with skeletal deformities (McKusick and Neufeld, 1983).  
           [0006]    There is a need, therefore, to purify α-N-acetylglucosaminidase and to clone genetic sequences encoding same to permit development of a range of therapeutic and diagnostic procedures to assist in the diagnosis and treatment of disease conditions arising from α-N-acetylglucosaminidase deficiency.  
         SUMMARY OF THE INVENTION  
         [0007]    One aspect of the invention provides an isolated nucleic acid molecule comprising a sequence of nucleotides which encodes or is complementary to a sequence which encodes a mammalian α-N-acetylglucosaminidase or fragment or derivative thereof.  
           [0008]    A second aspect of the invention provides an isolated nucleic acid molecule comprising a sequence of nucleotides which is capable of hybridising under at least low stringency conditions to a nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or a complementary strand or a homologue, analogue or derivative thereof.  
           [0009]    Another aspect of the invention is directed an isolated nucleic acid molecule which is at least 40% identical to the nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or to a complementary strand thereof or a homologue, analogue or derivative thereof.  
           [0010]    A further aspect of the present invention provides a nucleic acid molecule comprising a sequence of nucleotides encoding or complementary to a sequence encoding a polypeptide capable of hydrolysing the terminal α-N-acetylglucosamine residues present at the non-reducing terminus of fragments of heparan sulphate and heparin residues and wherein said nucleotide sequence is capable of hybridising under low stringency conditions to the nucleotide sequence set forth in SEQ ID NO:1.  
           [0011]    A further aspect of the invention is directed to a genetic construct comprising a sense molecule, for the expression or over-expression of α-N-acetylglucosaminidase in prokaryotic or eukaryotic cells.  
           [0012]    A further aspect of the present invention is directed to synthetic α-N-acetylglucosaminidase or like molecule.  
           [0013]    A further aspect of the invention contemplates antibodies to α-N-acetylglucosaminidase and preferably synthetic α-N-acetylglucosaminidase or a like molecule.  
           [0014]    In still yet another aspect of the present invention there is contemplated a method of diagnosing a mutation or other abberations in the α-N-acetylglucosaminidase gene in a human or animal patient.  
           [0015]    Another aspect contemplates a method of treating patients suffering from α-N-acetylglucosaminidase deficiency, such as in MPS-IIIB, said method comprising administering to said patient an effective amount of α-N-acetylglucosaminidase or active like form thereof.  
           [0016]    Another aspect of the present invention is directed to a pharmaceutical composition comprising a recombinant mammalian α-N-acetylglucosaminidase or an active fragment or derivative thereof and one or more pharmaceutically acceptable carriers and/or diluents. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0017]    [0017]FIG. 1 is a photographic representation of α-N-acetylglucosaminidase purified from human placenta following SDS/PAGE. Lane 1: M r  standards (kDa); Lanes 2 and 3: purified α-N-acetylglucosaminidase from human placenta. Lane 4 and 5, bovine serum albumin.  
         [0018]    [0018]FIG. 2 is a photographic representation of an SDS/polyacrylamide gel showing the molecular weights of recombinant α-N-acetylglucosaminidase polypeptides produced in CHO cells before (−) and after (+) PNGase F digestion. The 50 mM NaCl and 75 mM NaCl fractions are indicated. Molecular weights of α-N-acetylglucosaminidase polypeptides are indicated on the left of the figure. Molecular weights of marker proteins are indicated on the right hand side of the figure (lane 5). 
     
    
       [0019]    Single and three letter abbreviations of conventional amino acid residues as used herein are defined in Table 1.  
         [0020]    Suitable amino acid substitutions referred to herein are defined in Table 2.  
         [0021]    Codes for non-conventional amino acid residues as used herein are defined in Table 3.  
                               TABLE 1                                       Three-letter   One-letter           Amino Acid   Abbreviation   Symbol                           Alanine   Ala   A           Arginine   Arg   R           Asparagine   Asn   N           Aspartic acid   Asp   D           Cysteine   Cys   C           Glutamine   Gln   Q           Glutamic acid   Glu   E           Glycine   Gly   G           Histidine   His   H           Isoleucine   Ile   I           Leucine   Leu   L           Lysine   Lys   K           Methionine   Met   M           Phenylalanine   Phe   F           Proline   Pro   P           Serine   Ser   S           Threonine   Thr   T           Tryptophan   Trp   W           Tyrosine   Tyr   Y           Valine   Val   V           Any residue   Xaa   X                      
 
         [0022]    [0022]                         TABLE 2                           Suitable residues for amino acid substitutions            Original Residue   Exemplary Substitutions               Ala   Ser       Arg   Lys       Asn   Gln; His       Asp   Glu       Cys   Ser       Gln   Asn       Glu   Asp       Gly   Pro       His   Asn; Gln       Ile   Leu; Val       Leu   Ile; Val       Lys   Arg; Gln; Glu       Met   Leu; Ile       Phe   Met; Leu; Tyr       Ser   Thr       Thr   Ser       Trp   Tyr       Tyr   Trp; Phe       Val   Ile; Leu                    
         [0023]    [0023]                           TABLE 3                                   Non-conventional               amino acid   Code                           α-aminobutyric acid   Abu           α-amino-α-methylbutyrate   Mgabu           aminocyclopropane-   Cpro           carboxylate               aminoisobutyric acid   Aib           aminonorbornyl-   Norb           carboxylate               cyclohexylalanine   Chexa           cyclopentylalanine   Cpen           D-alanine   Dal           D-arginine   Darg           D-aspartic acid   Dasp           D-cysteine   Dcys           D-glutamine   Dgln           D-glutamic acid   Dglu           D-histidine   Dhis           D-isoleucine   Dile           D-leucine   Dleu           D-lysine   Dlys           D-methionine   Dmet           D-ornithine   Dorn           D-phenylalanine   Dphe           D-proline   Dpro           D-serine   Dser           D-threonine   Dthr           D-tryptophan   Dtrp           D-tyrosine   Dtyr           D-valine   Dval           D-α-methylalanine   Dmala           D-α-methylarginine   Dmarg           D-α-methylasparagine   Dmasn           D-α-methylaspartate   Dmasp           D-α-methylcysteine   Dmcys           D-α-methylglutamine   Dmgln           D-α-methylhistidine   Dmhis           D-α-methylisoleucine   Dmile           D-α-methylleucine   Dmleu           D-α-methyllysine   Dmlys           D-α-methylmethionine   Dmmet           D-α-methylornithine   Dmorn           D-α-methylphenylalanine   Dmphe           D-α-methylproline   Dmpro           D-α-methylserine   Dmser           D-α-methylthreonine   Dmthr           D-α-methyltryptophan   Dmtrp           D-α-methyltyrosine   Dmty           D-α-methylvaline   Dmval           D-N-methylalanine   Dnmala           D-N-methylarginine   Dnmarg           D-N-methylasparagine   Dnmasn           D-N-methylaspartate   Dnmasp           D-N-methylcysteine   Dnmcys           D-N-methylglutamine   Dnmgln           D-N-methylglutamate   Dnmglu           D-N-methylhistidine   Dnmhis           D-N-methylisoleucine   Dnmile           D-N-methylleucine   Dnmleu           D-N-methyllysine   Dnmlys           N-methylcyclohexylalanine   Nmchexa           D-N-methylornithine   Dnmorn           N-methylglycine   Nala           N-methylaminoisobutyrate   Nmaib           N-(1-methylpropyl)glycine   Nile           N-(2-methylpropyl)glycine   Nleu           D-N-methyltryptophan   Dnmtrp           D-N-methyltyrosine   Dnmtyr           D-N-methylvaline   Dnmval           γ-aminobutyric acid   Gabu           L-t-butylglycine   Tbug           L-ethylglycine   Etg           L-homophenylalanine   Hphe           L-α-methylarginine   Marg           L-α-methylaspartate   Masp           L-α-methylcysteine   Mcys           L-α-methylglutamine   Mgln           L-α-methylhistidine   Mhis           L-α-methylisoleucine   Mile           L-α-methylleucine   Mleu           L-α-methylmethionine   Mmet           L-α-methylnorvaline   Mnva           L-α-methylphenylalanine   Mphe           L-α-methylserine   Mser           L-α-methyltryptophan   Mtrp           L-α-methylvaline   Mval           N-(N-(2,2-diphenylethyl)   Nnbhm           carbamylmethyl)glycine               1-carboxy-1-(2,2-diphenyl-   Nmbc           ethylamino)cyclopropane           L-N-methylalanine   Nmala           L-N-methylarginine   Nmarg           L-N-methylasparagine   Nmasn           L-N-methylaspartic acid   Nmasp           L-N-methylcysteine   Nmcys           L-N-methylglutamine   Nmgln           L-N-methylglutamic acid   Nmglu           L-N-methylhistidine   Nmhis           L-N-methylisolleucine   Nmile           L-N-methylleucine   Nmleu           L-N-methyllysine   Nmlys           L-N-methylmethionine   Nmmet           L-N-methylnorleucine   Nmnle           L-N-methylnorvaline   Nmnva           L-N-methylornithine   Nmorn           L-N-methylphenylalanine   Nmphe           L-N-methylproline   Nmpro           L-N-methylserine   Nmser           L-N-methylthreonine   Nmthr           L-N-methyltryptophan   Nmtrp           L-N-methyltyrosine   Nmtyr           L-N-methylvaline   Nmval           L-N-methylethylglycine   Nmetg           L-N-methyl-t-butylglycine   Nmtbug           L-norleucine   Nle           L-norvaline   Nva           α-methyl-aminoisobutyrate   Maib           α-methyl-γ-aminobutyrate   Mgabu           α-methylcyclohexylalanine   Mchexa           α-methylcylcopentylalanine   Mcpen           α-methyl-α-napthylalanine   Manap           α-methylpenicillamine   Mpen           N-(4-aminobutyl)glycine   Nglu           N-(2-aminoethyl)glycine   Naeg           N-(3-aminopropyl)glycine   Norn           N-amino-α-methylbutyrate   Nmaabu           α-napthylalanine   Anap           N-benzylglycine   Nphe           N-(2-carbamylethyl)glycine   Ngln           N-(carbamylmethyl)glycine   Nasn           N-(2-carboxyethyl)glycine   Nglu           N-(carboxymethyl)glycine   Nasp           N-cyclobutylglycine   Ncbut           N-cycloheptylglycine   Nchep           N-cyclohexylglycine   Nchex           N-cyclodecylglycine   Ncdec           N-cylcododecylglycine   Ncdod           N-cyclooctylglycine   Ncoct           N-cyclopropylglycine   Ncpro           N-cycloundecylglycine   Ncund           N-(2,2-diphenylethyl)   Nbhm           glycine           N-(3,3-diphenylpropyl)   Nbhe           glycine           N-(3-guanidinopropyl)   Narg           glycine           N-(1-hydroxyethyl)glycine   Nthr           N-(hydroxyethyl))glycine   Nser           N-(imidazolylethyl))   Nhis           glycine           N-(3-indolylyethyl)   Nhtrp           glycine           N-methyl-γ-aminobutyrate   Nmgabu           D-N-methylmethionine   Dnmmet           N-methylcyclopentylalanine   Nmcpen           D-N-methylphenylalanine   Dnmphe           D-N-methylproline   Dnmpro           D-N-methylserine   Dnmser           D-N-methylthreonine   Dnmthr           N-(1-methylethyl)glycine   Nval           N-methyla-napthylalanine   Nmanap           N-methylpenicillamine   Nmpen           N-(p-hydroxyphenyl)glycine   Nhtyr           N-(thiomethyl)glycine   Ncys           penicillamine   Pen           L-α-methylalanine   Mala           L-α-methylasparagine   Masn           L-α-methyl-t-butylglycine   Mtbug           L-methylethylglycine   Metg           L-α-methylglutamate   Mglu           L-α-methylhomo   Mhphe           phenylalanine           N-(2-methylthioethyl)   Nmet           glycine           L-α-methyllysine   Mlys           L-α-methylnorleucine   Mnle           L-α-methylornithine   Morn           L-α-methylproline   Mpro           L-α-methylthreonine   Mthr           L-α-methyltyrosine   Mtyr           L-N-methylhomo   Nmhphe           phenylalanine           N-(N-(3,3-diphenylpropyl)   Nnbhe           carbamylmethyl)glycine                        
       DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
       [0024]    The present invention provides an isolated nucleic acid molecule comprising a sequence of nucleotides which encodes, or are complementary to a sequence which encodes, a mammalian α-N-acetylglucosaminidase or fragment or derivative thereof or its like molecule.  
         [0025]    Preferably, the mammal is a human, livestock animal, companion animal, wild animal or laboratory test animal (e.g. rabbit, rat, mouse or guinea pig). Most preferably, the mammal is a human. Conveniently, the α-N-acetylglucosaminidase is isolatable from the liver, kidney or placenta. However, the present invention extends to all mammalian α-N-acetylglucosaminidase enzymes and from any anatomical or cellular source and/or any biological fluid source, such as but not limited to plasma, serum, cell extract or lymph fluid.  
         [0026]    Although a preferred embodiment of the present invention contemplates the use of human α-N-acetylglucosaminidase or genomic or recombinant (e.g. cDNA) genetic sequences encoding same in the investigation, diagnosis and/or treatment of human subjects (i.e. homologous system), one skilled in the art will appreciate that the enzyme or genetic sequences encoding same from a non-human animal may also be useful. Such a heterologous system is encompassed by the present invention.  
         [0027]    The term “nucleic acid molecule” as used herein shall be taken to refer to any RNA or DNA (eg. cDNA) molecule, whether single-stranded or double-stranded or in a linear or covalently-closed form. The nucleic acid molecule may also be DNA corresponding to the entire genomic gene or a substantial portion thereof or a fragment or derivative thereof.  
         [0028]    The nucleic acid molecule of the present invention may constitute solely the nucleotide sequence encoding α-N-acetylglucosaminidase or a α-N-acetylglucosaminidase-like molecule or may be part of a larger nucleic acid molecule. Accordingly, the present invention extends to the isolated genomic α-N-acetylglucosaminidase gene. The non-translated sequences in a larger nucleic acid molecule may include vector, transcriptional and/or translational regulatory sequences, promoter, terminator, enhancer, replication or signal sequences or non-coding regions (eg intron sequences) of an isolated genomic gene.  
         [0029]    Reference herein to a “gene” is to be taken in its broadest context and includes:  
         [0030]    (i) a classical genomic gene consisting of transcriptional and/or translational regulatory sequences and/or a coding region and/or non-translated sequences (i.e. introns, 5′- and 3′-untranslated sequences);  
         [0031]    (ii) mRNA or cDNA corresponding to the coding regions (i.e. exons) optionally comprising 5′- or 3′-untranslated sequences of the gene; or  
         [0032]    (iii) synthetic, amplified DNA fragments or other recombinant nucleic acid molecules produced in vitro and comprising all or a part of the coding region and/or 5′- or 3′-untranslated sequences of the gene.  
         [0033]    The term “gene” is also used to describe synthetic or fusion molecules encoding all or part of a functional product. A functional product is one which comprises a sequence of nucleotides or is complementary to a sequence of nucleotides which encodes a functional polypeptide, in particular a polypeptide having the catalytic activity of α-N-acetylglucosaminidase or a homologue, analogue or derivative thereof.  
         [0034]    For the present purpose, “homologues” of a nucleotide sequence shall be taken to refer to an isolated nucleic acid molecule which is substantially the same as the nucleic acid molecule of the present invention or its complementary nucleotide sequence, notwithstanding the occurrence within said sequence, of one or more nucleotide substitutions, insertions, deletions, or rearrangements.  
         [0035]    “Analogues” of a nucleotide sequence set forth herein shall be taken to refer to an isolated nucleic acid molecule which is substantially the same as a nucleic acid molecule of the present invention or its complementary nucleotide sequence, notwithstanding the occurrence of any non-nucleotide constituents not normally present in said isolated nucleic acid molecule, for example carbohydrates, radiochemicals including radionucleotides, reporter molecules such as, but not limited to DIG, alkaline phosphatase or horseradish peroxidase, amongst others.  
         [0036]    “Derivatives” of a nucleotide sequence set forth herein shall be taken to refer to any isolated nucleic acid molecule which contains significant sequence similarity to said sequence or a part thereof. Generally, the nucleotide sequence of the present invention may be subjected to mutagenesis to produce single or multiple nucleotide substitutions, deletions and/or insertions. Nucleotide insertional derivatives of the nucleotide sequence of the present invention include 5′ and 3′ terminal fusions as well as intra-sequence insertions of single or multiple nucleotides or nucleotide analogues. Insertional nucleotide sequence variants are those in which one or more nucleotides or nucleotide analogues are introduced into a predetermined site in the nucleotide sequence of said sequence, although random insertion is also possible with suitable screening of the resulting product being performed. Deletional variants are characterised by the removal of one or more nucleotides from the nucleotide sequence. Substitutional nucleotide variants are those in which at least one nucleotide in the sequence has been removed and a different nucleotide or nucleotide analogue inserted in its place.  
         [0037]    Preferably, a homologue, analogue or derivative of an α-N-acetylglucosaminidase gene according to any embodiments described herein, comprises a sequence of nucleotides of at least 10 contiguous nucleotides derived from SEQ ID NO:1 or SEQ ID NO:3 or a complementary strand thereof, wherein the sequence of said homologue, analogue or derivative is at least 40% identical to SEQ ID NO:1 or SEQ ID NO:3 or a complementary strand thereof or wherein said homologue, analogue or derivative is capable of hybridising to said sequence under at least low stringency hybridisation conditions.  
         [0038]    For the purposes of nomenclature, the nucleotide sequence set for in SEQ ID NO: 1 relates to the cDNA encoding the human α-N-acetylglucosaminidase enzyme.  
         [0039]    The nucleotide sequence set forth in SEQ ID NO:3 relates to the genomic gene equivalent of the cDNA encoding the human liver α-N-acetylglucosaminidase enzyme. Those skilled in the art will be aware that the specific exon sequences described in SEQ ID NO:3 correspond to the coding regions of the α-N-acetylglucosaminidase gene, said exon regions further comprising the entire open reading frame of the cDNA sequence set forth in SEQ ID NO:1, when aligned in a head-to-tail configuration. The intron sequences of SEQ ID NO:3, which correspond to non-coding regions of the gene which are spliced from the primary transcription product thereof, although not explicitly defined, may be readily deduced by those skilled in the art, when provided with the exon sequence data provided in the nucleotide sequence listing.  
         [0040]    The nucleotide sequence of the present invention may correspond to the sequence of the naturally-occurring α-N-acetylglucosaminidase gene or may comprise a homologue, analogue or derivative thereof which contains single or multiple nucleotide substitutions, deletions and/or additions. All such homologues, analogue or derivatives encode α-N-acetylglucosaminidase or α-N-acetylglucosaminidase-like molecules or a homologue, analogue or derivative thereof as contemplated by the present invention. The length of the nucleotide sequence may-vary-from a few bases, such as-in nucleic acid probes or primers, to a full length sequence.  
         [0041]    The present invention is particularly directed to the nucleic acid in cDNA form and particularly when inserted into an, expression vector. The expression vector may be replicable in a eukaryotic or prokaryotic cell and may either produce mRNA or the mRNA may be subsequently translated into α-N-acetylglucosaminidase or like molecule. Particularly preferred eukaryotic cells include CHO cells but may be in any other suitable mammalian cells or cell lines or non-mammalian cells such as yeast or insect cells.  
         [0042]    In an alternative embodiment, the present invention provides a nucleic acid molecule comprising a sequence of nucleotides which encodes or are complementary to a sequence which encodes a polypeptide capable of hydrolysing the α-N-acetylglucosamine residues from the non-reducing terminus of heparan sulphate and heparin fragments and wherein said nucleotide sequence is capable of hybridising under at least low stringency conditions to a nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or a homologue, analogue or derivative thereof.  
         [0043]    A second aspect of the invention provides an isolated nucleic acid molecule comprising a sequence of nucleotides which is capable of hybridising under at least low stringency conditions to a nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or a complementary strand or a homologue, analogue or derivative thereof.  
         [0044]    Preferably, hybridisation is possible under at least medium stringent conditions. More preferably, hybridisation is possible under high stringent conditions.  
         [0045]    For the purposes of defining the level of stringency, reference can conveniently be made to Sambrook et al (1989) or Ausubel et al (1987) which are herein incorporated by reference.  
         [0046]    A low stringency is defined herein as being a hybridisation and/or wash carried out in 4-6× SSC/0.1-0.5% w/v SDS at 37-45° C. for 2-3 hours. A medium stringency hybridisation and/or wash is carried out in 1-4× SSC/0.25-0.5% w/v SDS at ≧45° C. for 2-3 hours and a high stringency hybridisation and/or wash is carried out 0.1-1× SSC/0.1% w/v SDS at 60° C. for 1-3 hours.  
         [0047]    Alternative conditions of stringency may be employed to those specifically recited herein. Generally, the stringency is increased by reducing the concentration of SSC buffer, and/or increasing the concentration of SDS and/or increasing the temperature of the hybridisation and/or wash. Those skilled in the art will be aware that the conditions for hybridisation and/or wash may vary depending upon the nature of the hybridisation membrane or the type of hybridisation probe used. Conditions for hybridisations and washes are well understood by one normally skilled in the art. For the purposes of clarification of parameters affecting hybridisation between nucleic acid molecules, reference is found in pages 2.10.8 to 2.10.16. of Ausubel et al. (1987), which is herein incorporated by reference.  
         [0048]    Those skilled in the art will be aware that the nucleotide sequences set forth in SEQ ID NO:1 and SEQ ID NO:3 may be used to isolate the corresponding genes from other human tissues or alternatively, from the tissues or cells of other species, without undue experimentation. Means for the isolated of such related sequences will also be known to those skilled in the art, for example nucleic acid hybridisation, polymerase chain reaction, antibody screening of expression libraries, functional screening of expression libraries, or complementation of mutants, amongst others. The present invention is not to be limited by the source from which the specific gene sequences described herein have been isolated or by the means used to isolate said sequences.  
         [0049]    In one embodiment, a related genetic sequence comprising genomic DNA, or mRNA, or cDNA is contacted with a hybridisation effective amount of a genetic sequence which encodes α-N-acetylglucosaminidase, or its complementary nucleotide sequence or a homologue, analogue, derivative or functional part thereof, and then said hybridisation is detected using a suitable detection means.  
         [0050]    The related genetic sequence may be in a recombinant form, in a virus particle, bacteriophage particle, yeast cell, animal cell, or a plant cell. Preferably, the related genetic sequence originates from an animal species or a human. More preferably, the related genetic sequence originates from a human.  
         [0051]    Preferably, the genetic sequence which encodes α-N-acetylglucosaminidase (i.e probe or latter genetic sequence) comprises a sequence of nucleotides of at least 10 nucleotides, more preferably at least 20 nucleotides, even more preferably at least 50 nucleotides and even still more preferably at least 100 nucleotides derived from the nucleotide sequence set forth in SEQ ID NO: 1 or SEQ ID NO:3 or a complementary sequence or a homologue, analogue or derivative thereof.  
         [0052]    Preferably, the detection means is a reporter molecule capable of giving an identifiable signal (e.g. a radioisotope such as  32 P or  35 S or a biotinylated molecule) covalently attached to the α-N-acetylglucosaminidase probe.  
         [0053]    In an alternative embodiment, the detection means is a polymerase chain reaction. According to this embodiment, two opposing non-complementary nucleic acid “primer molecules” of at least 10 nucleotides in length, more preferably at least 20 nucleotides in length, derived from the nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 may be contacted with a nucleic acid “template molecule” and specific nucleic acid molecule copies of the template molecule amplified in a polymerase chain reaction.  
         [0054]    The opposing primer molecules are selected such that they are each capable of hybridising to complementary strands of the same template molecule, wherein DNA polymerase-dependant DNA synthesis occurring from a first opposing primer molecule will be in a direction toward the second opposing primer molecule.  
         [0055]    Accordingly, both primers hybridise to said template molecule such that, in the presence of a DNA polymerase enzyme, a cofactor and appropriate substrate, DNA synthesis occurs in the 5′ to 3′ direction from each primer molecule towards the position on the DNA where the other primer molecule is hybridised, thereby amplifying the intervening DNA.  
         [0056]    Those skilled in the art are aware of the technical requirements of the polymerase chain reaction and are capable of any modifications which may be made to the reaction conditions. For example, of the polymerase chain reaction may be used in any suitable format, such as amplified fragment length polymorphism (AFLP), single-strand chain polymorphism (SSCP), amplification and mismatch detection (AMD), interspersed repetitive sequence polymerase chain reaction (IRS-PCR), inverse polymerase chain reaction (iPCR) and reverse transcription polymerase chain reaction (RT-PCR), amongst others, to isolate a related α-N-acetylglucosaminidase gene sequence or identify a mutation in an α-N-acetylglucosaminidase genetic sequence. Such variations of the polymerase chain reaction are discussed in detail by McPherson et al (1991), which is incorporated herein by reference. The present invention encompasses all such variations, the only requirement being that the final product of the reaction is an isolated nucleic acid molecule which is capable of encoding α-N-acetylglucosaminidase or a homologue, analogue or derivative thereof.  
         [0057]    In a preferred embodiment, the first primer molecule is preferably derived from the sense strand of a gene which encodes α-N-acetylglucosaminidase, in particular from the nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or a homologue, derivative or analogue thereof and the second primer molecule is preferably derived from the antisense strand of said gene.  
         [0058]    Those skilled in the art will be aware that it is not essential to the performance of the invention that the primer molecules be derived from the same gene.  
         [0059]    According to this embodiment, the nucleic acid primer molecule may further consist of a combination of any of the nucleotides adenine, cytidine, guanine, thymidine, or inosine, or functional analogues or derivatives thereof, capable of being incorporated into a polynucleotide molecule provided that it is capable of hybridising under at least low stringency conditions to the nucleic acid molecule set forth in SEQ ID NO:1 or SEQ ID NO:3 or a homologue, analogue or derivative thereof.  
         [0060]    The nucleic acid primer molecules may further be each contained in an aqueous pool comprising other nucleic acid primer molecules. More preferably, the nucleic acid primer molecule is in a substantially pure form.  
         [0061]    The nucleic acid template molecule may be in a recombinant form, in a virus particle, bacteriophage particle, yeast cell, animal cell, or a plant cell. Preferably, the related genetic sequence originates from a cell, tissue, or organ derived from an animal species or a human. More preferably, the related genetic sequence originates from a cell, tissue, or organ derived from a human.  
         [0062]    Accordingly, a third aspect of the present invention extends to an isolated nucleic acid molecule which is at least 40% identical to the nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or to a complementary strand thereof or a homologue, analogue or derivative thereof.  
         [0063]    Preferably, the percentage identity to SEQ ID NO:1 or SEQ ID NO:3 is at least about 55%, still more preferably at least about 65%, yet still more preferably at least about 75-80% and even still more preferably at least about 85-95%.  
         [0064]    In an even more preferred embodiment, the present invention provides an isolated nucleic acid molecule which is at least 40% identical to the nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3 or to a complementary strand thereof or a homologue, analogue or derivative thereof and is capable of hybridising under at least low stringency conditions to a nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:3.  
         [0065]    In a particularly preferred embodiment, the isolated nucleic acid molecule described herein is further capable of encoding a sequence of amino acids which is capable of carrying out the enzyme reaction catalysed by a α-N-acetylglucosaminidase enzyme.  
         [0066]    The isolated nucleic acid molecule of the present invention is also useful for developing a genetic construct comprising a sense molecule, for the expression or over-expression of α-N-acetylglucosaminidase in prokaryotic or eukaryotic cells. Particularly preferred eukaryotic cells include CHO cells but may be in any other suitable mammalian cells or cell lines or non-mammalian cells such as yeast or insect cells.  
         [0067]    The term “sense molecule” as used herein shall be taken to refer to an isolated nucleic acid molecule of the invention as described herein, which is provided in a format suitable for its expression to produce a recombinant polypeptide, when said sense molecule is introduced into a host cell.  
         [0068]    In a particularly preferred embodiment, a sense molecule which encodes the α-N-acetylglucosaminidase comprises a sequence of nucleotides set forth in SEQ ID NO:1 or SEQ ID NO:3 or a complementary strand, homologue, analogue or derivative thereof.  
         [0069]    In a most particularly preferred embodiment, the sense molecule of the invention comprises the sequence of nucleotides set forth in SEQ ID NO:1 or a complementary strand, homologue, analogue or derivative thereof.  
         [0070]    Those skilled in the art will be aware that expression of a sense molecule may require the nucleic acid molecule of the invention to be placed in operable connection with a promoter sequence to produce a “sense construct”. The choice of promoter for the present purpose may vary depending upon the level of expression of the sense molecule required and/or the tissue-specificity or developmental-specificity of expression of the sense molecule which is required. The sense construct may further comprise a terminator sequence and be introduced into a suitable host cell where it is capable of being expressed to produce a recombinant polypeptide gene product.  
         [0071]    In the context of the present invention, a sense molecule which corresponds to a genetic sequence or isolated nucleic acid molecule which encodes α-N-acetylglucosaminidase polypeptide or a homologue, analogue or derivative thereof, placed operably under the control of a suitable promoter sequence, is introduced into a cell using any suitable method for the transformation of said cell and said genetic sequence or isolated nucleic acid molecule is expressed therein to produce said polypeptide.  
         [0072]    The present invention clearly extends to genetic constructs designed to facilitate expression of any nucleic acid molecule described herein.  
         [0073]    A genetic construct of the present invention comprises the foregoing sense molecule, placed operably under the control of a promoter sequence capable of regulating the expression of the said nucleic acid molecule in a prokaryotic or eukaryotic cell, preferably a mammalian cell such as a CHO cell, a yeast cell, insect cell or bacterial cell. The said genetic construct optionally comprises, in addition to a promoter and sense molecule, a terminator sequence.  
         [0074]    The term “terminator” refers to a DNA sequence at the end of a transcriptional unit which signals termination of transcription. Terminators are 3′-non-translated DNA sequences containing a polyadenylation signal, which facilitates the addition of polyadenylate sequences to the 3′-end of a primary transcript. Terminators active in plant cells are known and described in the literature. They may be isolated from bacteria, fungi, viruses, animals and/or plants.  
         [0075]    Reference herein to a “promoter” is to be taken in its broadest context and includes the transcriptional regulatory sequences of a classical genomic gene, including the TATA box which is required for accurate transcription initiation, with or without a CCAAT box sequence and additional regulatory elements (i.e. upstream activating sequences, enhancers and silencers) which alter gene expression in response to developmental and/or external stimuli, or in a tissue-specific manner. A promoter is usually, but not necessarily, positioned upstream or 5′, of a structural gene, the expression of which it regulates. Furthermore, the regulatory elements comprising a promoter are usually positioned within 2 kb of the start site of transcription of the gene.  
         [0076]    In the present context, the term “promoter” is also used to describe a synthetic or fusion molecule, or derivative which confers, activates or enhances expression of said sense molecule in a cell.  
         [0077]    Preferred promoters may contain additional copies of one or more specific regulatory elements, to further enhance expression of the sense molecule and/or to alter the spatial expression and/or temporal expression of said sense molecule. For example, regulatory elements which confer copper inducibility may be placed adjacent to a heterologous promoter sequence driving expression of a sense molecule, thereby conferring copper inducibility on the expression of said molecule.  
         [0078]    Placing a sense molecule under the regulatory control of a promoter sequence means positioning the said molecule such that expression is controlled by the promoter sequence. Promoters are generally positioned 5′ (upstream) to the genes that they control. In the construction of heterologous promoter/structural gene combinations it is generally preferred to position the promoter at a distance from the gene transcription start site that is approximately the same as the distance between that promoter and the gene it controls in its natural setting, i.e., the gene from which the promoter is derived. As is known in the art, some variation in this distance can be accommodated without loss of promoter function. Similarly, the preferred positioning of a regulatory sequence element with respect to a heterologous gene to be placed under its control is defined by the positioning of the element in its natural setting, i.e., the genes from which it is derived. Again, as is known in the art, some variation in this distance can also occur.  
         [0079]    Examples of promoters suitable for use in genetic constructs of the present invention include viral, fungal, bacterial, animal and plant derived promoters capable of functioning in animal, human, yeast, insect or bacterial cells. The promoter may regulate the expression of the said molecule constitutively, or differentially with respect to the tissue in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, or plant pathogens, or metal ions, amongst others. Preferably, the promoter is capable of regulating expression of a sense-molecule in a cell derived from an animal species or human.  
         [0080]    In a particularly preferred embodiment, the promoter is derived from the genomic gene encoding α-N-acetylglucosaminidase, preferably the human α-N-acetylglucosaminidase gene. In a more preferred embodiment, however, the promoter is derived from the nucleotide sequence set forth in SEQ ID NO:3 or is at least capable of hybridising to nucleotide residues 1 to 989 of SEQ ID NO:3 or at least 20 contiguous nucleotides derived therefrom.  
         [0081]    In an even more particularly preferred embodiment, the promoter is the CMV promoter sequence or a promoter sequence derived therefrom.  
         [0082]    An alternative embodiment of the invention is directed to a genetic construct comprising a promoter or functional derivative, part fragment, homologue, or analogue thereof, derived from the α-N-acetylglucosaminidase genomic gene defined by SEQ ID NO: 3.  
         [0083]    Preferably, said genetic construct further comprises the α-N-acetylglucosaminidase sequence defined by SEQ ID NO:1 placed in operably connection with said promoter.  
         [0084]    A further aspect of the present invention is directed to synthetic α-N-acetylglucosaminidase or like molecule.  
         [0085]    The term “synthetic” as used herein shall be taken to include both recombinant and chemically-synthesised molecules produced by the sequential addition of amino acid residues or groups of amino acid residues in defined order.  
         [0086]    In one embodiment, the invention relates to recombinant α-N-acetylglucosaminidase or like molecule encoded by or expressed from the nucleic acid molecules as hereinbefore described.  
         [0087]    In another embodiment the synthetic α-N-acetylglucosaminidase or like molecule comprises a sequence of amino acids which is at least 40% identical to the amino acid sequence set forth in any one of SEQ ID Nos:2, 4, 5 or 6.  
         [0088]    More preferably, the percentage identity is at least 60% and still more preferably at least 80% or 85-90%.  
         [0089]    A particularly preferred embodiment of the present invention provides a synthetic α-N-acetylglucosaminidase as hereinbefore defined which comprises a sequence of amino acids substantially as set forth in any one of SEQ ID Nos:2, 4, 5 or 6 or a homologue, analogue or derivative thereof.  
         [0090]    For the purposes of nomenclature, the amino acid sequence set forth in SEQ ID NO:2 comprises the full-length translation product of the human α-N-acetylglucosaminidase gene (i.e. hereinafter referred to as the “α-N-acetylglucosaminidase polypeptide” or “SEQ ID NO:2”) produced by expression of either the cDNA sequence defined by SEQ ID NO:1 or the genomic gene defined by SEQ ID NO:3. The α-N-acetylglucosaminidase polypeptide comprises at least seven potentially-glycosylated Asn residues, at positions 261, 272, 435, 503, 513, 526 and 532. Furthermore, the amino acid sequence of the α-N-acetylglucosaminidase polypeptide may comprise a signal peptide of approximately 23 amino acid residues in length, with a probable site for signal peptide peptidase cleavage occurring between Gly 23  and Asp 24 .  
         [0091]    The amino acid sequences set forth in SEQ ID Nos:4-6 relate to N-terminal and internal (i.e. CNBr) amino acid sequences derived from human α-N-acetylglucosaminidase, purified as described in Example 1. As described in Example 2, the purified form of the enzyme comprises two polypeptides having approximate molecular weights of 82 and 77 kDa. The sequence set forth in SEQ ID NO:4 relates to the N-terminal sequence of the 82 kDa polypeptide, while SEQ ID NO:5 relates to the N-terminal sequence of the 77 kDa polypeptide. Furthermore, SEQ ID NO:4 comprises amino acids residues 24-43 of SEQ ID NO:2, while SEQ ID NO:5 comprises amino acid residues 59-76 of SEQ ID NO:2.  
         [0092]    The amino acid sequence defined by SEQ ID NO:6 relates to the CNBr-cleaved peptide of purified human α-N-acetylglucosaminidase. This amino acid sequence aligns with amino acid residues 540-554 of the α-N-acetylglucosaminidase polypeptide (SEQ ID NO:2).  
         [0093]    In the present context, “homologues” of a polypeptide refer to those polypeptides, enzymes or proteins which have a similar α-N-acetylglucosaminidase enzyme activity, notwithstanding any amino acid substitutions, additions or deletions thereto. A homologue may be isolated or derived from the same or another animal species.  
         [0094]    Furthermore, the amino acids of a homologous polypeptide may be replaced by other amino acids having similar properties, for example hydrophobicity, hydrophilicity, hydrophobic moment, charge or antigenicity, and so on.  
         [0095]    “Analogues” encompass α-N-acetylglucosaminidase polypeptides and peptide derivatives thereof notwithstanding the occurrence of any non-naturally occurring amino acid analogues therein.  
         [0096]    The term “derivative” in relation to the polypeptides of the invention refer to mutants, parts or fragments of a functional molecule. Derivatives include modified peptides in which ligands are attached to one or more of the amino acid residues contained therein, such as carbohydrates, enzymes, proteins, polypeptides or reporter molecules such as radionuclides or fluorescent compounds. Glycosylated, fluorescent, acylated or alkylated forms of the subject peptides are particularly contemplated by the present invention. Additionally, derivatives of a polypeptide may comprise fragments or parts of an amino acid sequence disclosed herein and are within the scope of the invention, as are homopolymers or heteropolymers comprising two or more copies of the subject polypeptides. Procedures for derivatizing peptides are well-known in the art.  
         [0097]    Accordingly, this aspect of the present invention is directed to any proteinaceous molecule comprising an amino acid sequence corresponding to the full length mammalian α-N-acetylglucosaminidase enzyme or to a like molecule. The like molecule, therefore, comprises parts, derivatives and/or portions of the α-N-acetylglucosaminidase enzyme whether functional or not.  
         [0098]    Preferably, the mammal is human but may be of non-human origin as contemplated above.  
         [0099]    The synthetic or recombinant α-N-acetylglucosaminidase of the present invention may comprise an amino acid sequence corresponding to the naturally occurring amino acid sequence or may contain single or multiple amino acid substitutions, deletions and/or additions. The length of the amino acid sequence may range from a few residues to a full length molecule.  
         [0100]    Amino acid substitutions are typically of single residues. Amino acid insertions will usually be in the order of about 1-10 amino acid residues and deletions will range from about 1-20 residues. Preferably, deletions or insertions are made in adjacent pairs, i.e. a deletion of two residues or insertion of two residues.  
         [0101]    Amino acid insertional derivatives of α-N-acetylglucosaminidase of the present invention include amino and/or carboxyl terminal fusions as well as intra-sequence insertions of single or multiple amino acids. Insertional amino acid sequence variants are those in which one or more amino acid residues are introduced into a predetermined site in the protein although random insertion is also possible with suitable screening of the resulting product. Deletional variants are characterised by the removal of one or more amino acids from the sequence. Substitutional amino acid variants are those in which at least one residue in the sequence has been removed and a different residue inserted in its place. Typical substitutions are those made in accordance with the following Table 2:  
         [0102]    The amino acid variants referred to above may readily be made using peptide synthetic techniques well known in the art such as solid phase peptide synthesis (Merrifield synthesis) and the like, or by recombinant DNA manipulations. Techniques for making substitution mutations at predetermined sites in DNA having known or partially known sequence are well known and include, for example, M13 mutagenesis. The manipulation of DNA sequence to produce variant proteins which manifest as substitutional, insertional or deletional variants are conveniently elsewhere described such as Sambrook et al, 1989  Molecular Cloning: A Laboratory Manual  Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y.  
         [0103]    The derivatives or like molecules include single or multiple substitutions, deletions and/or additions of any component(s) naturally or artificially associated with the α-N-acetylglucosaminidase enzyme such as carbohydrate, lipid and/or other proteinaceous moieties. For example, the present invention extends to glycosylated and non-glycosylated forms of the molecule. All such molecules are encompassed by the expression “mutants”, “derivatives”, “fragments”, “portions” and “like” molecules. These molecules may be active or non-active and may contain specific regions, such as a catalytic region. Particularly, preferred derivative molecules include those with altered glycosylation patterns relative to the naturally occurring molecule. Even more particularly, the recombinant molecule is more highly glycosylated than the naturally occurring molecule. Such highly glycosylated derivatives may have improved take-up properties and enhanced half-lives.  
         [0104]    As indicated in the Examples, the molecular weight of purified human α-N-acetylglucosaminidase (i.e. 82 kDa and 77 kDa) and recombinant mammalian α-N-acetylglucosaminidase produced in CHO cells (i.e. 89 kDa and 79 kDa) are greater than the deduced molecular weight of the α-N-acetylglucosaminidase polypeptide set forth in SEQ ID No:2 (i.e. 70 kDa), suggesting that the purified and recombinant polypeptide are post-translationally modified. The data presented in Example 8 indicate further that the recombinant α-N-acetylglucosaminidase enzyme produced in CHO cells, at least, is glycosylated and that the difference in molecular weight determined for the recombinant polypeptides and the polypeptide of SEQ ID No: 2 is due almost entirely to glycosylation of the recombinant polypeptide by CHO cells. As shown in Example 9, the glycosylated recombinant α-N-acetylglucosaminidase polypeptide exhibits enzymatic activity.  
         [0105]    The present invention also extends to synthetic α-N-acetylglucosaminidase or like molecules when fused to other proteinaceous molecules. The latter may include another enzyme, reporter molecule, purification site or an amino acid sequence which facilitates transport of the molecule out of a cell, such as a signal sequence.  
         [0106]    The present invention extends further to post-translational modifications to the α-N-acetylglucosaminidase enzyme. The modifications may be made to the naturally occurring enzyme or following synthesis by recombinant techniques. The modifications may be at the structural level or at, for example, the electrochemical level such as modifying net charge or structural conformation of the enzyme.  
         [0107]    Such modification may be important to facilitate entry or penetration of the enzyme into selected tissues such as cartilage or blood brain barriers or to increase circulation half-life.  
         [0108]    Analogues of α-N-acetylglucosaminidase contemplated herein include, but are not limited to, modifications to side chains, incorporation of unnatural amino acids and/or their derivatives during peptide synthesis and the use of crosslinkers and other methods which impose conformational constraints on the enzyme.  
         [0109]    Examples of side chain modifications contemplated by the present invention include modifications of amino groups such as by reductive alkylation by reaction with an aldehyde followed by reduction with NaBH 4 ; amidination with methylacetimidate; acylation with acetic anhydride; carbamoylation of amino groups with cyanate; trinitrobenzylation of amino groups with 2,4,6-trinitrobenzene sulphonic acid (TNBS); acylation of amino groups with succinic anhydride and tetrahydrophthalic anhydride; and pyridoxylation of lysine with pyridoxal-5′-phosphate followed by reduction with NaBH 4 .  
         [0110]    The guanidino group of arginine residues may be modified by the formation of heterocyclic condensation products with reagents such as 2,3-butanedione, phenylglyoxal and glyoxal.  
         [0111]    The carboxyl group may be modified by carbodiimide activation via O-acylisourea formation followed by subsequent derivatisation, for example, to a corresponding amide.  
         [0112]    Sulphydryl groups may be modified by methods such as carboxymethylation with iodoacetic acid or iodoacetamide; performic acid oxidation to cysteic acid; formation of a mixed disulphides with other thiol compounds; reaction with maleimide, maleic anhydride or other substituted maleimide; formation of mercurial derivatives using 4-chloromercuribenzoate, 4-chloromercuriphenylsulphonic acid, phenylmercury chloride, 2-chloromercuric-4-nitrophenol and other mercurials; carbamoylation with cyanate at alkaline pH.  
         [0113]    Tryptophan residues may be modified by, for example, oxidation with N-bromosuccinimide or alkylation of the indole ring with 2-hydroxy-5-nitrobenzyl bromide or sulphenyl halides. Tyrosine residues on the other hand, may be altered by nitration with tetranitromethane to form a 3-nitrotyrosine derivative.  
         [0114]    Modification of the imidazole ring of a histidine residue may be accomplished by alkylation with iodoacetic acid derivatives or N-carbethoxylation with diethylpyrocarbonate.  
         [0115]    Examples of incorporating unnatural amino acids and derivatives during peptide synthesis include, but are not limited to, use of norleucine, 4-amino butyric acid, 4-amino-3-hydroxy-5-phenylpentanoic acid, 6-aminohexanoic acid, t-butylglycine, norvaline, phenylglycine, ornithine, sarcosine, 4-amino-3-hydroxy-6-methylheptanoic acid, 2-thienyl alanine and/or D-isomers of amino acids. Non-naturally occurring amino acids contemplated by the present invention are incorporated herein, as Table 3.  
         [0116]    Crosslinkers can be used, for example, to stabilise 3D conformations, using homo-bifunctional crosslinkers such as the bifunctional imido esters having (CH 2 ) n  spacer groups with n=1 to n=6, glutaraldehyde, N-hydroxysuccinimide esters and hetero-bifunctional reagents which usually contain an amino-reactive moiety such as N-hydroxysuccinimide and another group specific-reactive moiety such as maleimido or dithio moiety (SH) or carbodiimide (COOH). In addition, the enzyme could be conformationally constrained by, for example, incorporation of C α  and N α -methylamino acids, introduction of double bonds between C α  and C β  atoms of amino acids and the formation of cyclic peptides or analogues by introducing covalent bonds such as forming an amide bond between the N and C termini, between two side chains or between a side chain and the N or C terminus.  
         [0117]    Electrochemical modifications of α-N-acetylglucosaminidase include interaction with polylysine or polyethylene glycol or other agent which effects an overall change to the net charge of the enzyme.  
         [0118]    Advantageously, the recombinant α-N-acetylglucosaminidase is a biologically pure preparation meaning that it has undergone some purification away for other proteins and/or non-proteinaceous material. The purity of the preparation may be represented as at least 40% of the enzyme, preferably at least 60%, more preferably at least 75%, even more preferably at least 85% and still more preferably at least 95% relative to non-α-N-acetylglucosaminidase material as determined by weight, activity, amino acid homology or similarity, antibody reactivity or other convenient means.  
         [0119]    Particularly preferred methods-for the preparation and purification of recombinant α-N-acetylglucosaminidase are provided in Examples 7 and 8.  
         [0120]    Those skilled in the art will be aware of the means of purifying a synthetic or recombinant α-N-acetylglucosaminidase from several sources without undue experimentation and for expressing the degree of purity of such a purified preparation of the enzyme.  
         [0121]    The present invention further contemplates antibodies to α-N-acetylglucosaminidase and preferably synthetic α-N-acetylglucosaminidase or like molecule. The antibodies may be polyclonal or monoclonal, naturally occurring or synthetic (including recombinant, fragment or fusion forms). Such antibodies will be useful in developing immunoassays for α-N-acetylglucosaminidase and for identifying additional genetic sequences which are capable of expressing α-N-acetylglucosaminidase polypeptides or homologues, analogues or derivatives thereof.  
         [0122]    Both polyclonal and monoclonal antibodies are obtainable by immunisation with an appropriate synthetic or recombinant gene product, or epitope, or peptide fragment of a gene product, in particular a α-N-acetylglucosaminidase polypeptide or a homologue, analogue or derivative thereof.  
         [0123]    Alternatively, fragments of antibodies may be used, such as Fab fragments. The present invention extends further to encompass recombinant and synthetic antibodies and to antibody hybrids. A “synthetic antibody” is considered herein to include fragments and hybrids of antibodies.  
         [0124]    A further aspect of the present invention contemplates a method of screening for mutations or other abberations in the α-N-acetylglucosaminidase gene in a human or animal patient. Such a method may be accomplished in a number of ways including isolating a source of DNA to be tested or mRNA therefrom and hybridising thereto a nucleic acid molecule as hereinbefore described. Generally, the nucleic acid is probe or primer size and polymerase chain reaction is a convenient means by which to analyse the RNA or DNA. Other suitable assays include the ligation chain reaction and the strand displacement amplification methods. The α-N-acetylglucosaminidase sequence can also be determined and compared to the naturally occurring sequence. Such methods may be useful in adults and children and may be adapted for a pre-natal test The DNA to be tested includes a genomic sample carrying the α-N-acetylglucosaminidase gene, a cDNA clone and/or amplification product.  
         [0125]    In accordance with this aspect of the present invention there is provided a method for screening for abberations in the α-N-acetylglucosaminidase gene including the absence of such a gene or a portion or a substantial portion thereof comprising isolating a sample of DNA or mRNA corresponding to a region of said DNA and contacting same with an oligonucleotide probe capable of hybridising to one or more complementary sequences within the α-N-acetylglucosaminidase gene and then detecting the hybridisation, the extent of hybridisation or the absence of hybridisation.  
         [0126]    Alternatively, the probe is a primer and capable of directing amplification of one or more regions of said α-N-acetylglucosaminidase gene and the amplification products and/or profile of amplification products is compared to an individual carrying the full gene or to a reference date base.  
         [0127]    Conveniently, the amplification products are sequenced to determine the presence or absence of the full gene.  
         [0128]    The present invention extends to the use of any and all DNA-based or nucleic acid-based hybridisation and/or polymerase chain reaction formats as described herein, for the diagnosis of a disorder involving the α-N-acetylglucosaminidase gene in a human or animal patient.  
         [0129]    The present invention further extends to a method of treating patients suffering from α-N-acetylglucosaminidase deficiency, such as in MPS-IIIB, said method comprising administering to said patient an effective amount of α-N-acetylglucosaminidase or active like form thereof.  
         [0130]    Preferably, the α-N-acetylglucosaminidase is in recombinant form. Such a method is referred to as “enzyme therapy”. Alternatively, gene therapy can be employed including introducing an active gene (i.e. a nucleic acid molecule as hereinbefore described) or to parts of the gene or other sequences which facilitate expression of a naturally occurring α-N-acetylglucosaminidase gene.  
         [0131]    Administration of α-N-acetylglucosaminidase for enzyme therapy may be by oral, intravenous, suppository, intraperitoneal, intramuscular, intranasal, intradermal or subcutaneous administration or by infusion or implantation. The α-N-acetylglucosaminidase is preferably as hereinbefore described including active mutants or derivatives thereof and glycosylation variants thereof. Administration may also be by way of gene therapy including expression of the gene by inclusion of the gene in viral vectors which are introduced into the animal (e.g. human) host to be treated. Alternatively, the gene may be expressed in a bacterial host which is then introduced and becomes part of the bacterial flora in the animal to be tested.  
         [0132]    Still yet another aspect of the present invention is directed to a pharmaceutical composition comprising synthetic (e.g. recombinant) α-N-acetylglucosaminidase or like molecule, including active derivatives and fragments thereof, alone or in combination with other active molecules. Such other molecules may act synergistically with the enzyme or facilitates its entry to a target cell. The composition will also contain one or more pharmaceutically acceptable carriers and/or diluents. The composition may alternatively comprise a genetic component useful in gene therapy.  
         [0133]    The active ingredients of the pharmaceutical composition comprising the synthetic or recombinant α-N-acetylglucosaminidase or mutants or fragments or derivatives thereof are contemplated to exhibit excellent activity in treating patients with a deficiency in the enzyme when administered in an amount which depends on the particular case. The variation depends, for example, on the patient and the α-N-acetylglucosaminidase used. For example, from about 0.5 ug to about 20 mg of enzyme per animal body or, depending on the animal and other factors, per kilogram of body weight may be administered. Dosage regima may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily, weekly, monthly or in other suitable time intervals or the dose may be proportionally reduced as indicated by the exigencies of the situation. Accordingly, alternative dosages in the order of 1.0 μg to 15 mg, 2.0 μg to 10 mg or 10 μg to 5 mg may be administered in a single or as part of multiple doses. The active compound may be administered in a convenient manner such as by the oral, intravenous (where water soluble), intramuscular, subcutaneous, intranasal, intradermal or suppository routes or implanting (eg using slow release molecules). Depending on the route of administration, the active ingredients which comprise a synthetic (e.g. recombinant) α-N-acetylglucosaminidase or fragments, derivatives or mutants thereof may be required to be coated in a material to protect same from the action of enzymes, acids and other natural conditions which may inactivate said ingredients. For example, the low lipophilicity of α-N-acetylglucosaminidase will allow it to be destroyed in the gastrointestinal tract by enzymes capable of cleaving peptide bonds and in the stomach by acid hydrolysis. In order to administer the vaccine by other than parenteral administration, the enzyme will be coated by, or administered with, a material to prevent its inactivation. For example, the enzyme may be administered in an adjuvant, co-administered with enzyme inhibitors or in liposomes. Adjuvant is used in its broadest sense and includes any immune stimulating compound such as interferon. Adjuvants contemplated herein include resorcinols, non-ionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether. Conveniently, the adjuvant is Freund&#39;s Complete or Incomplete Adjuvant. Enzyme inhibitors include pancreatic trypsin inhibitor, diisopropylfluorophosphate (DEP) and trasylol. Liposomes include water-in-oil-in-water emulsions as well as conventional liposomes.  
         [0134]    The active compound may also be administered in dispersions prepared in glycerol, liquid polyethylene glycols, and/or mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.  
         [0135]    The pharmaceutical forms suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of superfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thirmerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.  
         [0136]    Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredient(s) into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield a powder of the active ingredient plus any additional desired ingredient from previously sterile-filtered solution thereof.  
         [0137]    When the α-N-acetylglucosaminidase of the present invention is suitably protected as described above, the composition may be orally administered, for example, with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsule, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet. For oral therapeutic administration, the active compound may be incorporated with excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 1% by weight of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 5 to about 80% of the weight of the unit. The amount of active compound in the vaccine compositions is such that a suitable dosage will be obtained. Preferred compositions or preparations according to the present invention are prepared, so that an oral dosage unit form contains between about 0.5 ug and 20 mg of active compound.  
         [0138]    The tablets, troches, pills, capsules and the like may also contain the following: a binder such as gum gragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such a sucrose, lactose or saccharin may be added or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavouring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar or both. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release reparations and formulations.  
         [0139]    As used herein “pharmaceutically acceptable carriers and/or diluents” include any and all solvents, dispersion media, aqueous solutions, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, use thereof in the pharmaceutical compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.  
         [0140]    The present invention further relates to the use of α-N-acetylglucosaminidase or active fragment, mutant or derivative thereof in the manufacture of a medicament for the treatment of patients suffering from a deficiency in the naturally occurring enzyme (e.g. MPS-IIIB).  
         [0141]    The present invention is further described with reference to the following non-limiting Examples.  
       EXAMPLE 1  
     Purification of α-N-acetylglucosaminidase  
       [0142]    α-N-acetylglucosaminidase was purified according to the method described in Weber et al. (1996). Enzyme was purified to homogeneity from human placenta. Evidence of purity is shown following SDS/PAGE which is represented in FIG. 1. All samples were reduced with dithiothreitol prior to electrophoresis.  
       EXAMPLE 2  
     Characterisation of α-N-acetylglucosaminidase  
       [0143]    Results presented in FIG. 1 show two polypeptides of about 82 kDa and 77 kDa molecular weight, which correspond to α-N-acetylglucosaminidase polypeptides purified from human placenta according to Example 1.  
       EXAMPLE 3  
     Amino Acid Sequence Determination  
       [0144]    The N-terminal amino acid sequences the 77 kDA and 82 kDa α-N-acetylglucosaminidase polypeptides, in addition to the amino acid sequence of an internal CNBr cleavage product of these peptides, were determined using the methods of Weber et al. (1996).  
         [0145]    The amino acid sequences are shown in Table 4.  
                             TABLE 4                       N-Terminal Amino Acid Sequences (SEQ ID NO:4 and       SEQ ID No:5) and CNBr peptide sequence       (SEQ ID No:6) determined from Purified Human       α-N-Acetylglucosaminidase                                    polypeptide 82 kDa   DEAREAAAVRALVARLLGPG                       polypeptide 77 kDa   KPGLDTYSLGGGGAAX 1  VR                       CNBr peptide   WRLLLTSAPSLX 1 TX 1 P                      
 
         [0146]    X 1  no residue could be identified for this position, indicating that this residue could be phosphorylated or glycosylated.  
       EXAMPLE 4  
     Cloning of α-N-acetylglucosaminidase cDNA  
       [0147]    Oligonucleotide probes were prepared based on the partial amino acid sequences obtained for the purified α-N-acetylglucosaminidase polypeptides (Example 3). The probes were subsequently used to screen a human peripheral blood leukocyte cDNA library. An approximately 2.6 kbp cDNA clone was isolated encoding most of the sequence of human α-N-acetylglucosaminidase (SEQ ID NO:1).  
         [0148]    The remaining α-N-acetylglucosaminidase coding sequence was obtained from the nucleotide sequence of the corresponding genomic gene (SEQ ID NO:3), isolated by hybridisation to a human chromosome 17 library (Weber et.al. 1996).  
         [0149]    The complete open reading frame is 2232 nucleotides long and encodes a 743 (plus stop codon) amino acid protein. The predicted molecular mass of the longest mature protein (minus the 23 amino acid N-terminal signal peptide) is about 79,622 daltons.  
         [0150]    The amino acid sequence of α-N-acetylglucosaminidase is shown in SEQ ID NO:2. The deduced molecular weight of the desired amino acid sequence of α-N-acetylglucosaminidase is approximately 70 kDa. The probable site of signal peptide peptidase cleavage is between amino acids 23 and 24. There are seven potential N-glycosylation sites in the sequence.  
         [0151]    The nucleotide sequence of the corresponding α-N-acetylglucosaminidase genomic gene (SEQ ID No:3) comprises 10380 bp including 889 bp of 5′ upstream sequence corresponding to at least at part of the α-N-acetylglucosaminidase promoter sequence, in addition to the nucleotide sequences of introns I, II, II, IV, V, in addition to 1326 bp of 3′-untranslated sequence.  
       EXAMPLE 5  
     Construction of an Expression Vector Comprising the α-N-acetylglucosaminidase cDNA Sequence  
       [0152]    The cDNA insert of 1 clone pbl 33, containing bases 107 to 2575 of the α-N-acetylglucosaminidase cDNA was excised with EcoRI and subcloned into pBluescript II SK-(Stratagene). A 178 bp XmaI fragment (bases 1 to 178 of the α-N-acetylglucosaminidase cDNA) from cosmid sub-clone 6.3, containing the start codon, was cloned into the pBluescript subclone to produce a full-length cDNA sequence in addition to 101 bp of 5′ non-translated sequence as well as 245 bp of 3′ non-translated region including the polyadenylation-site, the polyA-tail and linkerDNA. The full length cDNA was directionally cloned into the pCDNA3 expressionvector (Invitrogen) via the EcoRI and BamHI sites.  
       EXAMPLE 6  
     Expression of Recombinant α-N-acetylglucosaminidase  
       [0153]    Chinese Hamster Ovary (CHO) cells were transfected with expressionvector using the DOTAP transfection reagent (Boehringer Mannheim) according to the manufacturers instructions. Cells were grown in Ham&#39;s F12 medium, 10% (v/v) fetal calf serum, penicillin and streptomycin sulfate at 100 μg/ml each. Cells were grown in nonselective medium for 48 h and then incubated in medium containing 750 μg/ml G418 sulfate (Geniticin) until resistant colonies emerged.  
         [0154]    Single cell clones were grown up and 26 of them were tested for expression of recombinant α-N-acetylglucosaminidase with a fluorogenic α-N-acetylglucosaminidase substrate. (i.e. N-acetylglucosamine α-linked to 4-methylumbelliferone)  
       EXAMPLE 7  
     Large Scale α-N-acetylglucosaminidase Production  
       [0155]    2 g of Cytodex 2 microcarrier beads were swollen in 250 ml of PBS for 3 h at 37° C. with three changes of PBS and then autoclaved for 15 min at 120° C. (wet cycle). The beads were then rinsed with sterile growth medium (Coons/DMEM, 10% v/v fetal calf serum, penicillin and streptomycin sulfate at 100 μg/ml each and 0.1% w/v Pluronic F68) and transferred into a Techne stirrer culture flask. The microcarrier beads were inoculated with seven confluent 175 flasks of the cell clone showing the highest expression of recombinant α-N-acetylglucosaminidase, Growth medium was added up to 200 ml and the culture incubated with a stirrer speed of 20 rpm to achieve an even distribution of cells on the beads. The cells were allowed to attach to the beads for 16 h at low speed then medium was added up to 500 ml and the stirrer speed increased to 30 rpm. After a growth phase of 48 to 72 h with daily aerating to allow gas exchange the beads were completely covered with cells and the medium was exchanged for production medium (Coons/DMEM, no fetal calf serum, penicillin and streptomycin sulfate at 100 μg/ml each, 0.1% w/v Pluronic F68 and 5 mM NH 4 Cl). The glucose concentration was monitored daily and the medium replaced, when glucose fell below 5 mM every 203 days. The harvested medium contained approximately 2 mg α-N-acetylglucosaminidase protein per dm 3  of production medium.  
       EXAMPLE 8  
     Purification of Recombinant α-N-acetylglucosaminidase  
       [0156]    Production medium was dialysed against 50 mM NaAc pH 5.5 and loaded onto a heparin-agarose column equilibrated in the same buffer. After washing with NaAc buffer and NaAc/50 mM NaCl the column was eluted with 75 mM NaCl in NaAc buffer. The eluate was dialysed against 20 mM Tris/HCl pH 7.5, loaded onto a DEAE Scphacel column, washed with 25 mM NaCl in 20 mM Tris/HCl and then eluted with 50 and 75 mM NaCl in 20 mM Tris/HCl respectively.  
         [0157]    SDS-PAGE of the two eluates showed two polypeptide bands associated with enzyme activity with apparent molecular weights of 79 and 89 kDa. The smaller α-N-acetylglucosaminidase was eluted predominantly in the 50 mM NaCl fraction whereas the 89 kDa α-N-acetylglucosaminidase polypeptide was enriched in the 75 mM NaCl fraction (FIG. 2).  
         [0158]    The difference in apparent molecular weight of the recombinant α-N-acetylglucosaminidase polypeptides is due to the presence of additional carbohydrate side chains, since a digest with PNGase F, which cleaves off N-glycosylation groups, reduced both the 79 kDa and 89 kDa polypeptides to the polypeptide band having an apparent molecular weight of about 70 kDa (FIG. 2), which corresponds to the approximate molecular weight deduced from primary amino acid sequence data (SEQ ID No:2).  
       EXAMPLE 9  
     Characteristics of Recombinant α-N-acetylglucosaminidase  
       [0159]    No differences were observed between the enzyme activities of the 79 and 89 kDa recombinant α-N-acetylglucosaminidase polypeptides produced in CHO cells according to Example 7 and 8. With the fluorogenic N-acetylglucosamine α-linked to 4-methylumbelliferone (4-MU) substrate, the enzyme has a pH-optimum of 4.6 with a k M  of 5.34 mM and a V max  of 3.97×10 6  pmol/min/mg. Towards a  3 H-labelled disaccharide substrate it should a pH-optimum of 4.1 with a k M  of 0.0166 mM and a V max  of 4.48×10 4  pmol/min/mg.  
       EXAMPLE 10  
     Mutational Analysis of Sanfilippo B Patients  
       [0160]    Genomic DNA is isolated from cultivated skin fibroblasts of patients by extraction with Phenol/Chloroform and used to amplify the eight exons and adjacent intronic sequences individually by PCR.  
         [0161]    Primer sequences used in the amplification reaction are readily determined from the nucleotide sequences of the α-N-acetylglucosaminidase cDNA and genomic clones. set forth in SEQ ID No:1 or SEQ ID No:3. Amplification conditions are also readily determined without undue experimentation. Procedures for the design of PCR primers and amplification conditions are described in detail, for example, by McPherson et al. (1991). Differences in the primary sequence can be identified by separating the PCR products on a polyacrylamide gel under non-denaturing conditions (SSCP gels). Base changes, insertions and deletions will lead to a different band pattern compared with the wildtype in most of the cases, which can be visualised either by autoradiography of the gel after labelling the DNA during the PCR or by staining unlabelled DNA in the gel with silver. PCR products which show a different band pattern are sequences to identify the change. Other patient samples can be tested for mutations and polymorphism that were found by hybridisation with wildtype- and mutation-specific oligonucleotides (ASO).  
         [0162]    Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.  
       REFERENCES  
       [0163]    1. Ausubel, F. M., Brent, R, Kingston, R E, Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K. (1987). In: Current Protocols in Molecular Biology. Wiley Interscience (ISBN 047150338).  
         [0164]    2. Hopwood J J (1989) In: “ Heparin: Chemical and Biological Properties, Clinical Applications ” (Lane D W and Lindahl U, eds.), 190-229, Edward Arnold, London.  
         [0165]    3. McKusick V and Neufeld E (1983) In: “ The Metabolic Basis of Inherited Disease ” (Stanbury J B, Wyngaarden J B, Fredrickson D S, Goldstein J L and Brown M S, eds), 5th Ed., 751-771, McGraw-Hill, New York.  
         [0166]    4. McPherson, M. J., Quirke, P. and Taylor, G. R, (1991) In: PCR A Practical Approach. Oxford University Press, Oxford. (ISBN 0-19-96322L-X).  
         [0167]    5. O&#39;Brien J S, (1972)  Proc. Natl. Acad. Sci. USA  69: 1720-1722.  
         [0168]    6. Sambrook, J., Fritsch, E., and Maniatis, T. (1989) In:“Molecular Cloning” a laboratory manual, Cold Spring Harbour.  
         [0169]    7. Von Figura, K, and Kresse K (1972)  Biochem Biophys. Res. Commun,  48: 262-269  
         [0170]    8. Weber B, Scott H, Blanch L, Clements P, Morris C P, Anson D, Hopwood J, (1996) Nature Genetics (submitted)  
 
     
       
       
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                                                SEQUENCE LISTING  
               
               
                   
               
               
                   
               
               
                 (1) GENERAL INFORMATION:  
               
               
                   
               
               
                    (iii) NUMBER OF SEQUENCES: 6  
               
               
                   
               
               
                   
               
               
                 (2) INFORMATION FOR SEQ ID NO: 1:  
               
               
                   
               
               
                      (i) SEQUENCE CHARACTERISTICS:  
               
               
                           (A) LENGTH: 2575 base pairs  
               
               
                           (B) TYPE: nucleic acid  
               
               
                           (C) STRANDEDNESS: single  
               
               
                           (D) TOPOLOGY: linear  
               
               
                   
               
               
                     (ii) MOLECULE TYPE: cDNA  
               
               
                   
               
               
                     (vi) ORIGINAL SOURCE:  
               
               
                           (A) ORGANISM: Homo sapiens  
               
               
                           (F) TISSUE TYPE: Peripheral Blood  
               
               
                           (G) CELL TYPE: Leukocyte  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: CDS  
               
               
                           (B) LOCATION: 102..2330  
               
               
                   
               
               
                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:  
               
               
                   
               
               
                 CCCGGGCTTA GCCTTCGGGT CCACGTGGCC GGAGGCCGGC AGCTGATTGG ACGCGGGCCG     60  
               
               
                   
               
               
                 CCCCACCCCC TGGCCGTCGC GGGACCCGCA GGACTGAGAC C ATG GAG GCG GTG        113  
               
               
                   
               
               
                                                               Met Glu Ala Val  
               
               
                                                                 1  
               
               
                   
               
               
                 GCG GTG GCC GCG GCG GTG GGG GTC CTT CTC CTG GCC GGG GCC GGG GGC      161  
               
               
                 Ala Val Ala Ala Ala Val Gly Val Leu Leu Leu Ala Gly Ala Gly Gly  
               
               
                   5                  10                  15                  20  
               
               
                   
               
               
                 GCG GCA GGC GAC GAG GCC CGG GAG GCG GCG GCC GTG CGG GCG CTC GTG      209  
               
               
                 Ala Ala Gly Asp Glu Ala Arg Glu Ala Ala Ala Val Arg Ala Leu Val  
               
               
                                  25                  30                  35  
               
               
                   
               
               
                 GCC CGG CTG CTG GGG CCA GGC CCC GCG GCC GAC TTC TCC GTG TCG GTG      257  
               
               
                 Ala Arg Leu Leu Gly Pro Gly Pro Ala Ala Asp Phe Ser Val Ser Val  
               
               
                              40                  45                  50  
               
               
                   
               
               
                 GAG CGC GCT CTG GCT GCC AAG CCG GGC TTG GAC ACC TAC AGC CTG GGC      305  
               
               
                 Glu Arg Ala Leu Ala Ala Lys Pro Gly Leu Asp Thr Tyr Ser Leu Gly  
               
               
                          55                  60                  65  
               
               
                   
               
               
                 GGC GGC GGC GCG GCG CGC GTG CGG GTG CGC GGC TCC ACG GGC GTG GCG      353  
               
               
                 Gly Gly Gly Ala Ala Arg Val Arg Val Arg Gly Ser Thr Gly Val Ala  
               
               
                      70                  75                  80  
               
               
                   
               
               
                 GCC GCC GCG GGG CTG CAC CGC TAC CTG CGC GAC TTC TGT GGC TGC CAC      401  
               
               
                 Ala Ala Ala Gly Leu His Arg Tyr Leu Arg Asp Phe Cys Gly Cys His  
               
               
                  85                  90                  95                 100  
               
               
                   
               
               
                 GTG GCC TGG TCC GGC TCT CAG CTG CGC CTG CCG CGG CCA CTG CCA GCC      449  
               
               
                 Val Ala Trp Ser Gly Ser Gln Leu Arg Leu Pro Arg Pro Leu Pro Ala  
               
               
                                 105                 110                 115  
               
               
                   
               
               
                 GTG CCG GGG GAG CTG ACC GAG GCC ACG CCC AAC AGG TAC CGC TAT TAC      497  
               
               
                 Val Pro Gly Glu Leu Thr Glu Ala Thr Pro Asn Arg Tyr Arg Tyr Tyr  
               
               
                             120                 125                 130  
               
               
                   
               
               
                 CAG AAT GTG TGC ACG CAA AGC TAC TCC TTC GTG TGG TGG GAC TGG GCC      545  
               
               
                 Gln Asn Val Cys Thr Gln Ser Tyr Ser Phe Val Trp Trp Asp Trp Ala  
               
               
                         135                 140                 145  
               
               
                   
               
               
                 CGC TGG GAG CGA GAG ATA GAC TGG ATG GCG CTG AAT GGC ATC AAC CTG      593  
               
               
                 Arg Trp Glu Arg Glu Ile Asp Trp Met Ala Leu Asn Gly Ile Asn Leu  
               
               
                     150                 155                 160  
               
               
                   
               
               
                 GCA CTG GCC TGG AGC GGC CAG GAG GCC ATC TGG CAG CGG GTG TAC CTG      641  
               
               
                 Ala Leu Ala Trp Ser Gly Gln Glu Ala Ile Trp Gln Arg Val Tyr Leu  
               
               
                 165                 170                 175                 180  
               
               
                   
               
               
                 GCC TTG GGC CTG ACC CAG GCA GAG ATC AAT GAG TTC TTT ACT GGT CCT      689  
               
               
                 Ala Leu Gly Leu Thr Gln Ala Glu Ile Asn Glu Phe Phe Thr Gly Pro  
               
               
                                 185                 190                 195  
               
               
                   
               
               
                 GCC TTC CTG GCC TGG GGG CGA ATG GGC AAC CTG CAC ACC TGG GAT GGC      737  
               
               
                 Ala Phe Leu Ala Trp Gly Arg Met Gly Asn Leu His Thr Trp Asp Gly  
               
               
                             200                 205                 210  
               
               
                   
               
               
                 CCC CTG CCC CCC TCC TGG CAC ATC AAG CAG CTT TAC CTG CAG CAC CGG      785  
               
               
                 Pro Leu Pro Pro Ser Trp His Ile Lys Gln Leu Tyr Leu Gln His Arg  
               
               
                         215                 220                 225  
               
               
                   
               
               
                 GTC CTG GAC CAG ATG CGC TCC TTC GGC ATG ACC CCA GTG CTG CCT GCA      833  
               
               
                 Val Leu Asp Gln Met Arg Ser Phe Gly Met Thr Pro Val Leu Pro Ala  
               
               
                     230                 235                 240  
               
               
                   
               
               
                 TTC GCG GGG CAT GTT CCC GAG GCT GTC ACC AGG GTG TTC CCT CAG GTC      881  
               
               
                 Phe Ala Gly His Val Pro Glu Ala Val Thr Arg Val Phe Pro Gln Val  
               
               
                 245                 250                 255                 260  
               
               
                   
               
               
                 AAT GTC ACG AAG ATG GGC AGT TGG GGC CAC TTT AAC TGT TCC TAC TCC      929  
               
               
                 Asn Val Thr Lys Met Gly Ser Trp Gly His Phe Asn Cys Ser Tyr Ser  
               
               
                                 265                 270                 275  
               
               
                   
               
               
                 TGC TCC TTC CTT CTG GCT CCG GAA GAC CCC ATA TTC CCC ATC ATC GGG      977  
               
               
                 Cys Ser Phe Leu Leu Ala Pro Glu Asp Pro Ile Phe Pro Ile Ile Gly  
               
               
                             280                 285                 290  
               
               
                   
               
               
                 AGC CTC TTC CTG CGA GAG CTG ATC AAA GAG TTT GGC ACA GAC CAC ATC     1025  
               
               
                 Ser Leu Phe Leu Arg Glu Leu Ile Lys Glu Phe Gly Thr Asp His Ile  
               
               
                         295                 300                 305  
               
               
                   
               
               
                 TAT GGG GCC GAC ACT TTC AAT GAG ATG CAG CCA CCT TCC TCA GAG CCC     1073  
               
               
                 Tyr Gly Ala Asp Thr Phe Asn Glu Met Gln Pro Pro Ser Ser Glu Pro  
               
               
                     310                 315                 320  
               
               
                   
               
               
                 TCC TAC CTT GCC GCA GCC ACC ACT GCC GTC TAT GAG GCC ATG ACT GCA     1121  
               
               
                 Ser Tyr Leu Ala Ala Ala Thr Thr Ala Val Tyr Glu Ala Met Thr Ala  
               
               
                 325                 330                 335                 340  
               
               
                   
               
               
                 GTG GAT ACT GAG GCT GTG TGG CTG CTC CAA GGC TGG CTC TTC CAG CAC     1169  
               
               
                 Val Asp Thr Glu Ala Val Trp Leu Leu Gln Gly Trp Leu Phe Gln His  
               
               
                                 345                 350                 355  
               
               
                   
               
               
                 CAG CCG CAG TTC TGG GGG CCC GCC CAG ATC AGG GCT GTG CTG GGA GCT     1217  
               
               
                 Gln Pro Gln Phe Trp Gly Pro Ala Gln Ile Arg Ala Val Leu Gly Ala  
               
               
                             360                 365                 370  
               
               
                   
               
               
                 GTG CCC CGT GGC CGC CTC CTG GTT CTG GAC CTG TTT GCT GAG AGC CAG     1265  
               
               
                 Val Pro Arg Gly Arg Leu Leu Val Leu Asp Leu Phe Ala Glu Ser Gln  
               
               
                         375                 380                 385  
               
               
                   
               
               
                 CCT GTG TAT ACC CGC ACT GCC TCC TTC CAG GGC CAG CCC TTC ATC TGG     1313  
               
               
                 Pro Val Tyr Thr Arg Thr Ala Ser Phe Gln Gly Gln Pro Phe Ile Trp  
               
               
                     390                 395                 400  
               
               
                   
               
               
                 TGC ATG CTG CAC AAC TTT GGG GGA AAC CAT GGT CTT TTT GGA GCC CTA     1361  
               
               
                 Cys Met Leu His Asn Phe Gly Gly Asn His Gly Leu Phe Gly Ala Leu  
               
               
                 405                 410                 415                 420  
               
               
                   
               
               
                 GAG GCT GTG AAC GGA GGC CCA GAA GCT GCC CGC CTC TTC CCC AAC TCC     1409  
               
               
                 Glu Ala Val Asn Gly Gly Pro Glu Ala Ala Arg Leu Phe Pro Asn Ser  
               
               
                                 425                 430                 435  
               
               
                   
               
               
                 ACC ATG GTA GGC ACG GGC ATG GCC CCC GAG GGC ATC AGC CAG AAC GAA     1457  
               
               
                 Thr Met Val Gly Thr Gly Met Ala Pro Glu Gly Ile Ser Gln Asn Glu  
               
               
                             440                 445                 450  
               
               
                   
               
               
                 GTG GTC TAT TCC CTC ATG GCT GAG CTG GGC TGG CGA AAG GAC CCA GTG     1505  
               
               
                 Val Val Tyr Ser Leu Met Ala Glu Leu Gly Trp Arg Lys Asp Pro Val  
               
               
                         455                 460                 465  
               
               
                   
               
               
                 CCA GAT TTG GCA GCC TGG GTG ACC AGC TTT GCC GCC CGG CGG TAT GGG     1553  
               
               
                 Pro Asp Leu Ala Ala Trp Val Thr Ser Phe Ala Ala Arg Arg Tyr Gly  
               
               
                     470                 475                 480  
               
               
                   
               
               
                 GTC TCC CAC CCG GAC GCA GGG GCA GCG TGG AGG CTA CTG CTC CGG AGT     1601  
               
               
                 Val Ser His Pro Asp Ala Gly Ala Ala Trp Arg Leu Leu Leu Arg Ser  
               
               
                 485                 490                 495                 500  
               
               
                   
               
               
                 GTG TAC AAC TGC TCC GGG GAG GCC TGC AGG GGC CAC AAT CGT AGC CCG     1649  
               
               
                 Val Tyr Asn Cys Ser Gly Glu Ala Cys Arg Gly His Asn Arg Ser Pro  
               
               
                                 505                 510                 515  
               
               
                   
               
               
                 CTG GTC AGG CGG CCG TCC CTA CAG ATG AAT ACC AGC ATC TGG TAC AAC     1697  
               
               
                 Leu Val Arg Arg Pro Ser Leu Gln Met Asn Thr Ser Ile Trp Tyr Asn  
               
               
                             520                 525                 530  
               
               
                   
               
               
                 CGA TCT GAT GTG TTT GAG GCC TGG CGG CTG CTG CTC ACA TCT GCT CCC     1745  
               
               
                 Arg Ser Asp Val Phe Glu Ala Trp Arg Leu Leu Leu Thr Ser Ala Pro  
               
               
                         535                 540                 545  
               
               
                   
               
               
                 TCC CTG GCC ACC AGC CCC GCC TTC CGC TAC GAC CTG CTG GAC CTC ACT     1793  
               
               
                 Ser Leu Ala Thr Ser Pro Ala Phe Arg Tyr Asp Leu Leu Asp Leu Thr  
               
               
                     550                 555                 560  
               
               
                   
               
               
                 CGG CAG GCA GTG CAG GAG CTG GTC AGC TTG TAC TAT GAG GAG GCA AGA     1841  
               
               
                 Arg Gln Ala Val Gln Glu Leu Val Ser Leu Tyr Tyr Glu Glu Ala Arg  
               
               
                 565                 570                 575                 580  
               
               
                   
               
               
                 AGC GCC TAC CTG AGC AAG GAG CTG GCC TCC CTG TTG AGG GCT GGA GGC     1889  
               
               
                 Ser Ala Tyr Leu Ser Lys Glu Leu Ala Ser Leu Leu Arg Ala Gly Gly  
               
               
                                 585                 590                 595  
               
               
                   
               
               
                 GTC CTG GCC TAT GAG CTG CTG CCG GCA CTG GAC GAG GTG CTG GCT AGT     1937  
               
               
                 Val Leu Ala Tyr Glu Leu Leu Pro Ala Leu Asp Glu Val Leu Ala Ser  
               
               
                             600                 605                 610  
               
               
                   
               
               
                 GAC AGC CGC TTC TTG CTG GGC AGC TGG CTA GAG CAG GCC CGA GCA GCG     1985  
               
               
                 Asp Ser Arg Phe Leu Leu Gly Ser Trp Leu Glu Gln Ala Arg Ala Ala  
               
               
                         615                 620                 625  
               
               
                   
               
               
                 GCA GTC AGT GAG GCC GAG GCC GAT TTC TAC GAG CAG AAC AGC CGC TAC     2033  
               
               
                 Ala Val Ser Glu Ala Glu Ala Asp Phe Tyr Glu Gln Asn Ser Arg Tyr  
               
               
                     630                 635                 640  
               
               
                   
               
               
                 CAG CTG ACC TTG TGG GGG CCA GAA GGC AAC ATC CTG GAC TAT GCC AAC     2081  
               
               
                 Gln Leu Thr Leu Trp Gly Pro Glu Gly Asn Ile Leu Asp Tyr Ala Asn  
               
               
                 645                 650                 655                 660  
               
               
                   
               
               
                 AAG CAG CTG GCG GGG TTG GTG GCC AAC TAC TAC ACC CCT CGC TGG CGG     2129  
               
               
                 Lys Gln Leu Ala Gly Leu Val Ala Asn Tyr Tyr Thr Pro Arg Trp Arg  
               
               
                                 665                 670                 675  
               
               
                   
               
               
                 CTT TTC CTG GAG GCG CTG GTT GAC AGT GTG GCC CAG GGC ATC CCT TTC     2177  
               
               
                 Leu Phe Leu Glu Ala Leu Val Asp Ser Val Ala Gln Gly Ile Pro Phe  
               
               
                             680                 685                 690  
               
               
                   
               
               
                 CAA CAG CAC CAG TTT GAC AAA AAT GTC TTC CAA CTG GAG CAG GCC TTC     2225  
               
               
                 Gln Gln His Gln Phe Asp Lys Asn Val Phe Gln Leu Glu Gln Ala Phe  
               
               
                         695                 700                 705  
               
               
                   
               
               
                 GTT CTC AGC AAG CAG AGG TAC CCC AGC CAG CCG CGA GGA GAC ACT GTG     2273  
               
               
                 Val Leu Ser Lys Gln Arg Tyr Pro Ser Gln Pro Arg Gly Asp Thr Val  
               
               
                     710                 715                 720  
               
               
                   
               
               
                 GAC CTG GCC AAG AAG ATC TTC CTC AAA TAT TAC CCC GGC TGG GTG GCC     2321  
               
               
                 Asp Leu Ala Lys Lys Ile Phe Leu Lys Tyr Tyr Pro Gly Trp Val Ala  
               
               
                 725                 730                 735                 740  
               
               
                   
               
               
                 GGC TCT TGG TGATAGATTC GCCACCACTG GGCCTTGTTT TCCGCTAATT             2370  
               
               
                 Gly Ser Trp  
               
               
                   
               
               
                 CCAGGGCAGA TTCCAGGGCC CAGAGCTGGA CAGACATCAC AGGATAACCC AGGCCTGG     2430  
               
               
                   
               
               
                 GGAGGCCCCA CGGCCTGCTG GTGGGGTCTG ACCTGGGGGG ATTGGAGGGA AATGACCT     2490  
               
               
                   
               
               
                 CCTCCACCAC CACCCAAAGT GTGGGATTAA AGTACTGTTT TCTTTCCACT TAAAAAAA     2550  
               
               
                   
               
               
                 AAAAAAGTCG AGCGGCCGCG AATTC                                         2575  
               
               
                   
               
               
                   
               
               
                 (2) INFORMATION FOR SEQ ID NO: 2:  
               
               
                   
               
               
                      (i) SEQUENCE CHARACTERISTICS:  
               
               
                           (A) LENGTH: 743 amino acids  
               
               
                           (B) TYPE: amino acid  
               
               
                           (D) TOPOLOGY: linear  
               
               
                   
               
               
                     (ii) MOLECULE TYPE: protein  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 261  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 272  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 435  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 503  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 513  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 526  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Potentially-glycosylated Asn site,  
               
               
                           (B) LOCATION: 532  
               
               
                   
               
               
                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:  
               
               
                   
               
               
                 Met Glu Ala Val Ala Val Ala Ala Ala Val Gly Val Leu Leu Leu Ala  
               
               
                   1               5                  10                  15  
               
               
                   
               
               
                 Gly Ala Gly Gly Ala Ala Gly Asp Glu Ala Arg Glu Ala Ala Ala Val  
               
               
                              20                  25                  30  
               
               
                   
               
               
                 Arg Ala Leu Val Ala Arg Leu Leu Gly Pro Gly Pro Ala Ala Asp Phe  
               
               
                          35                  40                  45  
               
               
                   
               
               
                 Ser Val Ser Val Glu Arg Ala Leu Ala Ala Lys Pro Gly Leu Asp Thr  
               
               
                      50                  55                  60  
               
               
                   
               
               
                 Tyr Ser Leu Gly Gly Gly Gly Ala Ala Arg Val Arg Val Arg Gly Ser  
               
               
                  65                  70                  75                  80  
               
               
                   
               
               
                 Thr Gly Val Ala Ala Ala Ala Gly Leu His Arg Tyr Leu Arg Asp Phe  
               
               
                                  85                  90                  95  
               
               
                   
               
               
                 Cys Gly Cys His Val Ala Trp Ser Gly Ser Gln Leu Arg Leu Pro Arg  
               
               
                             100                 105                 110  
               
               
                   
               
               
                 Pro Leu Pro Ala Val Pro Gly Glu Leu Thr Glu Ala Thr Pro Asn Arg  
               
               
                         115                 120                 125  
               
               
                   
               
               
                 Tyr Arg Tyr Tyr Gln Asn Val Cys Thr Gln Ser Tyr Ser Phe Val Trp  
               
               
                     130                 135                 140  
               
               
                   
               
               
                 Trp Asp Trp Ala Arg Trp Glu Arg Glu Ile Asp Trp Met Ala Leu Asn  
               
               
                 145                 150                 155                 160  
               
               
                   
               
               
                 Gly Ile Asn Leu Ala Leu Ala Trp Ser Gly Gln Glu Ala Ile Trp Gln  
               
               
                                 165                 170                 175  
               
               
                   
               
               
                 Arg Val Tyr Leu Ala Leu Gly Leu Thr Gln Ala Glu Ile Asn Glu Phe  
               
               
                             180                 185                 190  
               
               
                   
               
               
                 Phe Thr Gly Pro Ala Phe Leu Ala Trp Gly Arg Met Gly Asn Leu His  
               
               
                         195                 200                 205  
               
               
                   
               
               
                 Thr Trp Asp Gly Pro Leu Pro Pro Ser Trp His Ile Lys Gln Leu Tyr  
               
               
                     210                 215                 220  
               
               
                   
               
               
                 Leu Gln His Arg Val Leu Asp Gln Met Arg Ser Phe Gly Met Thr Pro  
               
               
                 225                 230                 235                 240  
               
               
                   
               
               
                 Val Leu Pro Ala Phe Ala Gly His Val Pro Glu Ala Val Thr Arg Val  
               
               
                                 245                 250                 255  
               
               
                   
               
               
                 Phe Pro Gln Val Asn Val Thr Lys Met Gly Ser Trp Gly His Phe Asn  
               
               
                             260                 265                 270  
               
               
                   
               
               
                 Cys Ser Tyr Ser Cys Ser Phe Leu Leu Ala Pro Glu Asp Pro Ile Phe  
               
               
                         275                 280                 285  
               
               
                   
               
               
                 Pro Ile Ile Gly Ser Leu Phe Leu Arg Glu Leu Ile Lys Glu Phe Gly  
               
               
                     290                 295                 300  
               
               
                   
               
               
                 Thr Asp His Ile Tyr Gly Ala Asp Thr Phe Asn Glu Met Gln Pro Pro  
               
               
                 305                 310                 315                 320  
               
               
                   
               
               
                 Ser Ser Glu Pro Ser Tyr Leu Ala Ala Ala Thr Thr Ala Val Tyr Glu  
               
               
                                 325                 330                 335  
               
               
                   
               
               
                 Ala Met Thr Ala Val Asp Thr Glu Ala Val Trp Leu Leu Gln Gly Trp  
               
               
                             340                 345                 350  
               
               
                   
               
               
                 Leu Phe Gln His Gln Pro Gln Phe Trp Gly Pro Ala Gln Ile Arg Ala  
               
               
                         355                 360                 365  
               
               
                   
               
               
                 Val Leu Gly Ala Val Pro Arg Gly Arg Leu Leu Val Leu Asp Leu Phe  
               
               
                     370                 375                 380  
               
               
                   
               
               
                 Ala Glu Ser Gln Pro Val Tyr Thr Arg Thr Ala Ser Phe Gln Gly Gln  
               
               
                 385                 390                 395                 400  
               
               
                   
               
               
                 Pro Phe Ile Trp Cys Met Leu His Asn Phe Gly Gly Asn His Gly Leu  
               
               
                                 405                 410                 415  
               
               
                   
               
               
                 Phe Gly Ala Leu Glu Ala Val Asn Gly Gly Pro Glu Ala Ala Arg Leu  
               
               
                             420                 425                 430  
               
               
                   
               
               
                 Phe Pro Asn Ser Thr Met Val Gly Thr Gly Met Ala Pro Glu Gly Ile  
               
               
                         435                 440                 445  
               
               
                   
               
               
                 Ser Gln Asn Glu Val Val Tyr Ser Leu Met Ala Glu Leu Gly Trp Arg  
               
               
                     450                 455                 460  
               
               
                   
               
               
                 Lys Asp Pro Val Pro Asp Leu Ala Ala Trp Val Thr Ser Phe Ala Ala  
               
               
                 465                 470                 475                 480  
               
               
                   
               
               
                 Arg Arg Tyr Gly Val Ser His Pro Asp Ala Gly Ala Ala Trp Arg Leu  
               
               
                                 485                 490                 495  
               
               
                   
               
               
                 Leu Leu Arg Ser Val Tyr Asn Cys Ser Gly Glu Ala Cys Arg Gly His  
               
               
                             500                 505                 510  
               
               
                   
               
               
                 Asn Arg Ser Pro Leu Val Arg Arg Pro Ser Leu Gln Met Asn Thr Ser  
               
               
                         515                 520                 525  
               
               
                   
               
               
                 Ile Trp Tyr Asn Arg Ser Asp Val Phe Glu Ala Trp Arg Leu Leu Leu  
               
               
                     530                 535                 540  
               
               
                   
               
               
                 Thr Ser Ala Pro Ser Leu Ala Thr Ser Pro Ala Phe Arg Tyr Asp Leu  
               
               
                 545                 550                 555                 560  
               
               
                   
               
               
                 Leu Asp Leu Thr Arg Gln Ala Val Gln Glu Leu Val Ser Leu Tyr Tyr  
               
               
                                 565                 570                 575  
               
               
                   
               
               
                 Glu Glu Ala Arg Ser Ala Tyr Leu Ser Lys Glu Leu Ala Ser Leu Leu  
               
               
                             580                 585                 590  
               
               
                   
               
               
                 Arg Ala Gly Gly Val Leu Ala Tyr Glu Leu Leu Pro Ala Leu Asp Glu  
               
               
                         595                 600                 605  
               
               
                   
               
               
                 Val Leu Ala Ser Asp Ser Arg Phe Leu Leu Gly Ser Trp Leu Glu Gln  
               
               
                     610                 615                 620  
               
               
                   
               
               
                 Ala Arg Ala Ala Ala Val Ser Glu Ala Glu Ala Asp Phe Tyr Glu Gln  
               
               
                 625                 630                 635                 640  
               
               
                   
               
               
                 Asn Ser Arg Tyr Gln Leu Thr Leu Trp Gly Pro Glu Gly Asn Ile Leu  
               
               
                                 645                 650                 655  
               
               
                   
               
               
                 Asp Tyr Ala Asn Lys Gln Leu Ala Gly Leu Val Ala Asn Tyr Tyr Thr  
               
               
                             660                 665                 670  
               
               
                   
               
               
                 Pro Arg Trp Arg Leu Phe Leu Glu Ala Leu Val Asp Ser Val Ala Gln  
               
               
                         675                 680                 685  
               
               
                   
               
               
                 Gly Ile Pro Phe Gln Gln His Gln Phe Asp Lys Asn Val Phe Gln Leu  
               
               
                     690                 695                 700  
               
               
                   
               
               
                 Glu Gln Ala Phe Val Leu Ser Lys Gln Arg Tyr Pro Ser Gln Pro Arg  
               
               
                 705                 710                 715                 720  
               
               
                   
               
               
                 Gly Asp Thr Val Asp Leu Ala Lys Lys Ile Phe Leu Lys Tyr Tyr Pro  
               
               
                                 725                 730                 735  
               
               
                   
               
               
                 Gly Trp Val Ala Gly Ser Trp  
               
               
                             740  
               
               
                   
               
               
                   
               
               
                 (2) INFORMATION FOR SEQ ID NO: 3:  
               
               
                   
               
               
                      (i) SEQUENCE CHARACTERISTICS:  
               
               
                           (A) LENGTH: 10380 base pairs  
               
               
                           (B) TYPE: nucleic acid  
               
               
                           (C) STRANDEDNESS: single  
               
               
                           (D) TOPOLOGY: linear  
               
               
                   
               
               
                     (ii) MOLECULE TYPE: DNA (genomic)  
               
               
                   
               
               
                     (vi) ORIGINAL SOURCE:  
               
               
                           (A) ORGANISM: Homo sapiens  
               
               
                   
               
               
                   (viii) POSITION IN GENOME:  
               
               
                           (A) CHROMOSOME/SEGMENT: Chromosome 17  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: exon 1  
               
               
                           (B) LOCATION: 990..1372  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: exon 2  
               
               
                           (B) LOCATION: 2115..2262  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: exon 3  
               
               
                           (B) LOCATION: 3056..3202  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: exon 4  
               
               
                           (B) LOCATION: 3387..3472  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: exon 5  
               
               
                           (B) LOCATION: 5667..5923  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: exon 6  
               
               
                           (B) LOCATION: 7745..8955  
               
               
                   
               
               
                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:  
               
               
                   
               
               
                 ATAATGAGCA GTGAGGACGA TCAGAGGTCA CCTTCCTGTC TTGGTTTTGG CAGGTTTTGA     60  
               
               
                   
               
               
                 CCAGTTTCTT TGCTGCATTC TGTTTTATCA GCGGGGTCTT GTGACCTTTT ATCTTGTGCT    120  
               
               
                   
               
               
                 GACCTCCTGT CTCATCCTGT GACGAAGGCC TAACCTCCTG GGAATTCAGC CCAGCAGGTC    180  
               
               
                   
               
               
                 TCTGCCTCAT TTTACCCAGC CCCTGTTCAA GATGGAGTCG CTCTGGTTGG AAACTTCTGA    240  
               
               
                   
               
               
                 CAAAATGACA GCTCCTGTTA TGTTGCTGCT GCTGCCGCCA ATGGACAGCC TTTAACGTGC    300  
               
               
                   
               
               
                 CCGCCAGCCC TGCTCCACCG CCGGCCTGGG CTCACATGGC CCCATCCCTC CTCGAACCTC    360  
               
               
                   
               
               
                 CTAGCCTGTT AGTTACTCAA ATCTGCAAGC TCTCTGCCTT CTCAGGGCCT TCAATAAATG    420  
               
               
                   
               
               
                 CATTTCTTCT GTCTGGAAGG CTCTTCCTTT CCCTCTTCTA GCCAATTCCT ATTCATCCCT    480  
               
               
                   
               
               
                 GAGTTTCAGA TTAAAAGTCA CTTCCTTTGG AAACCTTACT TCGCTACTTC GCTACTTACT    540  
               
               
                   
               
               
                 GCACTACTTC GCAGCATCAC AACTATGATG GAAATCCTTA CTTACGTTAA ATATCTGGTT    600  
               
               
                   
               
               
                 TCTAGGTCAC CTCCCTGACG GGGACGGTAG GGACCGTCTT CTCGTTCATC AGTAGGGAAG    660  
               
               
                   
               
               
                 TAGCTATGGC AGTGCCTGAT ACAAAATAAA CTCCAAATGT GTATTTATTA GATGGTTGGA    720  
               
               
                   
               
               
                 TGGAAGTTAT TTGCGTGTGA AAGCGCGTTT TACCCGAAGG CGCTCTGTGA GGGCCAGCGG    780  
               
               
                   
               
               
                 GTCCCCTTCG GCCCTGGAGC CGGGGTCACA CGCTCCCCAC CGCGTGCGGT CACGAGACGC    840  
               
               
                   
               
               
                 CCCCAAGGGA GTATCCTGGT ACCCGGAAGC CGCGACTCCT GGCCCTGAGC CCGGGCTTAG    900  
               
               
                   
               
               
                 CCTTCGGGTC CACGTGGCCG GAGCCGGCAG CTGATTGGAC GCGGGCCGCC CCACCCCCTG    960  
               
               
                   
               
               
                 GCCGTCGCGG GACCCGCAGG ACTGAGACCA TGGAGGCGGT GGCGGTGGCC GCGGCGGTGG   1020  
               
               
                   
               
               
                 GGGTCCTTCT CCTGGCCGGG GCCGGGGGCG CGGCAGGCGA CGAGGCCCGG GAGGCGGCGG   1080  
               
               
                   
               
               
                 CCGTGCGGGC GCTCGTGGCC CGGCTGCTGG GGCCAGGCCC CGCGGCCGAC TTCTCCGTGT   1140  
               
               
                   
               
               
                 CGGTGGAGCG CGCTCTGGCT GCCAAGCCGG GCTTGGACAC CTACAGCCTG GGCGGCGGCG   1200  
               
               
                   
               
               
                 GCGCGGCGCG CGTGCGGGTG CGCGGCTCCA CGGGCGTGGC GGCCGCCGCG GGGCTGCACC   1260  
               
               
                   
               
               
                 GCTACCTGCG CGACTTCTGT GGCTGCCACG TGGCCTGGTC CGGCTCTCAG CTGCGCCTGC   1320  
               
               
                   
               
               
                 CGCGGCCACT GCCAGCCGTG CCGGGGGAGC TGACCGAGGC CACGCCCAAC AGGTACCGCC   1380  
               
               
                   
               
               
                 CCGAAGCTTC CCCGCGTCCG CCCGAGGCGC TTACCCCCTC CCGGAGCCGC TGCCACCCAA   1440  
               
               
                   
               
               
                 ATCGGGAGGC TGAGCGGGGA GCGCTGGCCG GAAGGCCCAG CTGCGCCGCC TCCAGCAGCT   1500  
               
               
                   
               
               
                 GTGTGGCCTT GAGCCAGCCA CTCTGCCTTT CAGAGCCTCG GCTGGCCCAC CTGAAAAACG   1560  
               
               
                   
               
               
                 GAAAGAAGAC GCCTACCGTG CAGTGTTATT GTGAGGATTT GCACGATGAT GGGCATAGAA   1620  
               
               
                   
               
               
                 TTTGTGGTGC ACAATTGGTG ATGAGTGAAT TTTCTTGCCT TCCTCCCCCA CCTTCTCTTT   1680  
               
               
                   
               
               
                 GAACCTGCGG ACTGAGGAAG GACGCCTCCA TCCCCCACCC TACAGGCCTG TGTTCCAGCG   1740  
               
               
                   
               
               
                 CCTGCCACAC TATGGAGTGA TGTGTTCACA CAGCTGTCCT CCCCTGCCCA TCTGTTAGAC   1800  
               
               
                   
               
               
                 TGTGGGGGCA GGGATTCCCC GTTCCAGGAA AACACCGTGC AGAGGAGGGG CTCTGGCAGT   1860  
               
               
                   
               
               
                 GTGGCATGAA AGTGGAATAT GCCACCCAAA TACCCGCCAG GCTAGAGGGC CCTGGGAGAG   1920  
               
               
                   
               
               
                 TGCAGGGGAC GAGTGCCTCA GAAGCCCAGC CCCGGTACCT GGTCTCAGCT CCACCTGGGG   1980  
               
               
                   
               
               
                 TGGGTCCCAG TGTGCAGCAG AAGGGCCGAG TTTGGAGCCC CTCCCCTCTC CTCTAGGTGG   2040  
               
               
                   
               
               
                 GGGATGGGGG ATTTGTTCCA GGGCCGTGGA CCCTCCAGGG TGGGATGCGC CCCTGCTCAT   2100  
               
               
                   
               
               
                 GACACTGCCC GCAGGTACCG CTATTACCAG AATGTGTGCA CGCAAAGCTA CTCCTTCGTG   2160  
               
               
                   
               
               
                 TGGTGGGACT GGGCCCGCTG GGAGCGAGAG ATAGACTGGA TGGCGCTGAA TGGCATCAAC   2220  
               
               
                   
               
               
                 CTGGCACTGG CCTGGAGCGG CCAGGAGGCC ATCTGGCAGC GGGTGCGTGC CCACTGTCCC   2280  
               
               
                   
               
               
                 TTCCCCACCC TCCTCTATGG CGGGAGCCAC CGTAGGTGTT TTCACCCGCC CCCCAGCATG   2340  
               
               
                   
               
               
                 GGCGCAGTGT CTCTCTCTAG AAGTGCTTTC AGCGTGCACA GTGGCTTGGG CCTCCTAAAA   2400  
               
               
                   
               
               
                 ACTGAGGCTT CCGGCCGGGC GCGGTGGCTC ACGCCTGTCA TCCCAGCACT TCGGGAGGCC   2460  
               
               
                   
               
               
                 TAGGCGGGCG GATCAGGAGT TCAGGAGATC GAGACCATCC TGGCCAACAT TGTGAAACCC   2520  
               
               
                   
               
               
                 CGTCTCTACT AAAATACAAA GAAATAGCAA CCTGGGCAAC AGAGCGAGAC TCTGTCTAAA   2580  
               
               
                   
               
               
                 AAAAAAAAAA AAAAAAACTG AGGCTTCCAG TTTGAGGAGT GGGGCTCCTT CCCCCATCTC   2640  
               
               
                   
               
               
                 CCCTATGCAG CCAATCACCT GGTCCCTTGG ATCCAACTCA TGGGCAGCTC TAGATCTGCC   2700  
               
               
                   
               
               
                 TCCCTGGAAG CTTCTGTGCT GCAATGGCTG CTCCAGGCTC TGCTTAAGCT CTTCACACAG   2760  
               
               
                   
               
               
                 TTGCCCTGCC CTTCCATCTG GCACTCTTGC TCCATGAAGC CTTCTAAGGC CTTCCTGTTG   2820  
               
               
                   
               
               
                 GGGGAAAGCC CCTTTGTGCC CCATCTCCTC ACCCATGCGA CAAAGGCAAC ACAGTGAACT   2880  
               
               
                   
               
               
                 CACCTACTCA CAGGTCTCTT TCCTCTGGGC TGTGGGCTCC TTGATGGCAG CGTTCGGATT   2940  
               
               
                   
               
               
                 TTGTCTCAGT AGCCCTAGCA CCCAGCACAA AGAAGCAATG AGTGAATGGT TGTTGAATGA   3000  
               
               
                   
               
               
                 ATGAATGAAT GAATGAAGAT GAATATATTT CTATGTGTGG GCCCTTCTTC CTCAGGTGTA   3060  
               
               
                   
               
               
                 CCTGGCCTTG GGCCTGACCC AGGCAGAGAT CAATGAGTTC TTTACTGGTC CTGCCTTCCT   3120  
               
               
                   
               
               
                 GGCCTGGGGG CGAATGGGCA ACCTGCACAC CTGGGATGGC CCCCTGCCCC CCTCCTGGCA   3180  
               
               
                   
               
               
                 CATCAAGCAG CTTTACCTGC AGGTAAAAGG ATGGAAAAGG GAAGGGGCAG AATCGGTGAT   3240  
               
               
                   
               
               
                 AGATGGTCAT GGGCCCAGGA AGGGTGGTAT TAGGCCGGCC CCAGGGCTCT TAACTGAGGC   3300  
               
               
                   
               
               
                 GGGGGGCTGC GTGTATCCTG GGAGATGAGG GCCTTCTCAT AGGACAGCAG TGGCCATGCT   3360  
               
               
                   
               
               
                 CACCACCCTT CCTTCTGTTC CTCCAGCACC GGGTCCTGGA CCAGATGCGC TCCTTCGGCA   3420  
               
               
                   
               
               
                 TGACCCCAGT GCTGCCTGCA TTCGCGGGGC ATGTTCCCGA GGCTGTCACC AGGTGAGGTT   3480  
               
               
                   
               
               
                 CCGCTCACCC CCTCCACTTA GCTCAGAGAG GGAATTTTAT TCCCTTCTAG AACATGACTT   3540  
               
               
                   
               
               
                 AAAAACTTAA GCTCTGGGCC GGGCGCAGTG GCTCACGCCT GTAATCCCAG CACTTTGGGA   3600  
               
               
                   
               
               
                 GGCCGAGTTG GGCGGATCAC CTGAGGTCAG GAGTTCGAGA CCAGCCTGGC CAACATGGTG   3660  
               
               
                   
               
               
                 AAACCCTGTC TCTACTAAAA ATATAAAAAT TAGCTGGGCA TGGTGGCACG CGCCTGTAAT   3720  
               
               
                   
               
               
                 CCCATCTACT TAGGAGGCTG AGACAGGAGA ATTGCTTAAA CCTGGGAGGC AGACGTTGCA   3780  
               
               
                   
               
               
                 GTGAGTCAAG ATCACGCCAT TGCACTCCAG CCTGGGTGAC GAGCGAAACT CTGTCTCAAA   3840  
               
               
                   
               
               
                 CAAACAAACA AGCTCTGGAC GTAGGCCTGG GTTTGATTTC TGACTCTGCT ACTAATTAGC   3900  
               
               
                   
               
               
                 TGTGTGACTT CGGGCAGATG ACATGACTGC TCTGTGCCTC AGTTTCCTTA CTTGTAAAAT   3960  
               
               
                   
               
               
                 GGGATCTCTA CCCACTTCGC TGTAGGGTTT GTAATTATCT CTCGATCTAT CTGTGACTTT   4020  
               
               
                   
               
               
                 GCACAGAGTG CTAGCAAATG GCAGCCCTTG GGAGTGGCAG CAGGGGTGCT CCAGTGTCCC   4080  
               
               
                   
               
               
                 TTGTCCCTCC TGTTCCTCTG TGCTTCCCAG CCATCCTCTC ACATGTGGTT GGGAAAAGTC   4140  
               
               
                   
               
               
                 TTCAAGGCTC ACCTGAGACC TCCCCTCCTT CAGGAAGCCT TGCTAGTGCC CCGCATGACC   4200  
               
               
                   
               
               
                 TCCTTTGCAC CTGCTAATGT CTGGCTCCCA TACTCTCGTA GGACTTAATG CATGCCAGTG   4260  
               
               
                   
               
               
                 GCCTCCCTGC CCGCCTCTTT GCCCCCATCA CCAGGTGGCA GGAAACTCAC TCATTCATTC   4320  
               
               
                   
               
               
                 AATAAACTTG GTCCAGCTGT CTGAGGCTGC CAGAACTGGC TGTGCTGGGT CCTGGGAGGC   4380  
               
               
                   
               
               
                 GGCAAGAAAG GTGCCCAAGG GCTTACCCCT GATAGGAGAG ATATGTTGGC TGAAGGATAC   4440  
               
               
                   
               
               
                 AATGTGGGGA CAAGGACAGG AATATATGTG GGTTCCGCTC TCCTCTGCCG GGAGAGAGGG   4500  
               
               
                   
               
               
                 GCAGGAAGGG CTCAGGGCAG AGCCCAGCCT TGAAAAATGA GTGTTGCTTG GACGGACGCT   4560  
               
               
                   
               
               
                 TGGCTAATGC TTGTAATCCT AGCGTTTTGG GAGGCTGAGG CGTATGGATC ACCTGCGGTC   4620  
               
               
                   
               
               
                 AGGAGTTAAA GACCAGCCTG GCCAACATGG CGAAACCCCA TCTCTACTAA AAGTACAAAA   4680  
               
               
                   
               
               
                 ATTAGCCAGG CGTGGTGGCG GGCTCCTGTA ATCCCAGCTA CTCGGTAGGC TGAGGCATGA   4740  
               
               
                   
               
               
                 GAATCTCTTG AAGCCAGGGG CCAGAGACTG CAGTGAGCCG AGATCACACC ACTTCACTCC   4800  
               
               
                   
               
               
                 AGCCTGGGTG ACAGAGTGAG ACTCCGTCTC AAAAAAAAAA AAAAAAAAAG GAAAGAAAAT   4860  
               
               
                   
               
               
                 TAAACACCTC ATGTTCTCAC TCATAGTGGG AGTTGAACAA TGAGAACAAC ATGGACACAG   4920  
               
               
                   
               
               
                 GAAGGGGAAC ATCACACACC GGGGCCTTTC GCGGTGTGGG GGTCAAGGGG AGGAGTAGCA   4980  
               
               
                   
               
               
                 TTGGGACAGA TACTTAATGC ATGCGGGGCT GAAAACCTAG ATGATGGGTT GATGGGTGCA   5040  
               
               
                   
               
               
                 GCAAACCACC ATGGCACATG TATACCTATG CAACAAACCT GCATGTTCTG CACAGAACTG   5100  
               
               
                   
               
               
                 AACTGAAAGT ATAATTAAAA AAAAAAAAAA AAGCTGGGTG CGGTGGCCCA CACCTGTAAT   5160  
               
               
                   
               
               
                 CCCAGCACTT TGGGAGGCCG AGACGGGCGG ATCACAAGGT CAGCAGATCG AGACCATCCT   5220  
               
               
                   
               
               
                 GGCTAACACA GTGAAACTCA GTCTCTACTA AAAATACAAA AAATTAGCCG GGTGTGGTGG   5280  
               
               
                   
               
               
                 CGGGCACCTG TAGTCCCAGC TACTAGGGAG GCTGAGGCAG GAGAATGGCA TGAACCTGGG   5340  
               
               
                   
               
               
                 AGGCAGAGCT TGCAGTGAGC TGAGAATGCG CCACTGCACT CCAGCCTGGG GGACAGAGTG   5400  
               
               
                   
               
               
                 AGACTCTGCC TCAAAAAAAA AAAAAAAAAG AAAGAAAAAG GAGCGTTGCT TGTTTCAGGC   5460  
               
               
                   
               
               
                 CACAGGAAGG GGAGAGATAG TGAAAGTTTT TCAGAGAAGG TGGCCAGGGA AGGAGAAGAA   5520  
               
               
                   
               
               
                 AGGACTGTAG GCAGAGAGCA TAGCCTGTAC AAAGCCATAG AGGCAAGAGA AACCAGGAGC   5580  
               
               
                   
               
               
                 TGTAGAGAAG TTGGCAAGGC TGTTGAACAC TATGGTGAAC ACTATGGCGG CTTCCATGAA   5640  
               
               
                   
               
               
                 ATATCTGAGC TTTTGCTCCC CACTAGGGTG TTCCCTCAGG TCAATGTCAC GAAGATGGGC   5700  
               
               
                   
               
               
                 AGTTGGGGCC ACTTTAACTG TTCCTACTCC TGCTCCTTCC TTCTGGCTCC GGAAGACCCC   5760  
               
               
                   
               
               
                 ATATTCCCCA TCATCGGGAG CCTCTTCCTG CGAGAGCTGA TCAAAGAGTT TGGCACAGAC   5820  
               
               
                   
               
               
                 CACATCTATG GGGCCGACAC TTTCAATGAG ATGCAGCCAC CTTCCTCAGA GCCCTCCTAC   5880  
               
               
                   
               
               
                 CTTGCCGCAG CCACCACTGC CGTCTATGAG GCCATGACTG CAGGTACAGT GCCTGGGTGG   5940  
               
               
                   
               
               
                 GGTGGGAGAG CCCCCCAGAC CCTCAAAAAG AAGGGAGTAG CAGATGTCAG TAGGGGTAGG   6000  
               
               
                   
               
               
                 CAGAGGGACT GGAATAATGC CTCGCCATAA CACACAGTAC TTTATAGTTT ACCAAGCACG   6060  
               
               
                   
               
               
                 TGTACACATG CGTTGTCTCA GTGAATCCCA CTGTGGTTGA GAGGTGAGCT CTGGAAGCCA   6120  
               
               
                   
               
               
                 ACAACCTGGG TCACACCTCG CGCTCCTATT TCCTGGCCGT GTGACTTATG ACTCATGACC   6180  
               
               
                   
               
               
                 TCCTTCCCAG TGTCTCGTTT GCTTTTCCTG TAAACTGGGA CTACCTCATA GGTAGAATAA   6240  
               
               
                   
               
               
                 CGCCTGGCCC AGAGCAAAGG CCACTAAGAG CTAGCTATGA ACAAGGATTT TGTTTCATCT   6300  
               
               
                   
               
               
                 CTGCGTGGTT GCTGAAGTAG GCACTGCAGG CAGGAGGTGA GTGGATGTGC CTAAAGGCAC   6360  
               
               
                   
               
               
                 TAAGTGCGCA TCCTGCTACA AAACTGTGAA GCCAGGGCTC CTTCCTGCCA CTTAAAGGAG   6420  
               
               
                   
               
               
                 GAGTGGAGCA GAGGGCGCCC AAGTCAGGAA TGACTTAGTG GAGAGGCGTC TGTGTTGGCC   6480  
               
               
                   
               
               
                 AGGAAGGGAA CAGATCAGCT CAGCCTTTCT TGAGCAGTAC TGCTCCAAGT GTGACCCAAA   6540  
               
               
                   
               
               
                 ACCAGCAGCA GCAGCAGCAG CAGCCCGAGC TGTGAGATGG CAAATTCTCA GGCCCTACCC   6600  
               
               
                   
               
               
                 AAGACCTGAA GGAGAAGCTA CATTTTTTTT TTTTTTGAGA CAGATTTCAC TCTGTTGCTG   6660  
               
               
                   
               
               
                 AGGCTGGAGC ACAGTGGCAC AATCTCATCT CACTGCAACC TTCGTCTCCT AGGTTCAAGC   6720  
               
               
                   
               
               
                 GATTCTCCTG CCTCAGCCTC CCGAGTAGCT GGGACTATAG GCACCCGCCA CCACGCCCGG   6780  
               
               
                   
               
               
                 CAATTTTTGT TTGTTTTGAG ATAGAGTCTC GCTCTGTCAC CCAGGCTGGA GTGCAGTGGC   6840  
               
               
                   
               
               
                 ACGATCTCAG TTCACTGCAA CCTCTGCTTC CTGAGTTCAA GCGATTCTCC TGCCTCAGCC   6900  
               
               
                   
               
               
                 TCCTGAGTAG CTGGGATTAC AGGCGCCCCC CAACCACACT CGGCTAATTT TTGTATTTTT   6960  
               
               
                   
               
               
                 AGTAGAGACG GGGTTTCGCT ATGTAGGTCA AGCTGGTTTC AAACTCCTGA CCTCAAATGA   7020  
               
               
                   
               
               
                 TTCGCCCACT TCAGCCTCCC AAAGTGCTGG GATTACAGGT GTGAGCCACC TTGCCTGGCC   7080  
               
               
                   
               
               
                 AATTTTTGTA TTTTTAGTAG AAACAGGTTT CACCATGGTG GCCAGACTGG TCTCAAACTC   7140  
               
               
                   
               
               
                 CTGACCTCAG GTGAACTGCC CACCTCAGCC TCCCAAAGTA CTGGTATTAC AGGCGTGATC   7200  
               
               
                   
               
               
                 CACTGCGACT GGCCTTGATT TTGTTTTTGA GACAGAATCT TACTCTGTCG CCCAGACTGG   7260  
               
               
                   
               
               
                 AGTGCAGTGG CACAATCTCA GCTCACTGCA ACTTCTGCCT CATGGGTTCA AGTGATTCTT   7320  
               
               
                   
               
               
                 GTGCCTCTAC CTCCCGAGTA GCCGGGATTA CAGGCACCTG CCATTACGCT AGGCTAATTT   7380  
               
               
                   
               
               
                 TTGTATTTTT AGTATAGACA GGGTTTCCCC ACATTGGCCA GGCTGGTCTG GAACTCCTGG   7440  
               
               
                   
               
               
                 GCTCAAGTGA TCCACCTGCT TCAGCCCCTC AGAGTACTGG GATTATAGGT GTGGGCCACC   7500  
               
               
                   
               
               
                 ACGCCCATTC AGAAACCTCC ATGTTTTAAG GAGCCCTCTG GGTAACTCTC ATGTTCACCC   7560  
               
               
                   
               
               
                 AAGCTGCTGA ACCCTGTCCT GGAGTTTTCA GAGGGACGCG TATGTGCCAC AGAGCGTCCC   7620  
               
               
                   
               
               
                 GCTGGTGGGG GTCATGGGAA GCCATGACCT GGGATAGACA GTCGTCTGTA GAGTGGGGTG   7680  
               
               
                   
               
               
                 AACATTCCCT GGGCCCTCTG TTTCATCACT CCTCTTCTCT GTTCCCCCTA CCTCCTGTCC   7740  
               
               
                   
               
               
                 ACAGTGGATA CTGAGGCTGT GTGGCTGCTC CAAGGCTGGC TCTTCCAGCA CCAGCCGCAG   7800  
               
               
                   
               
               
                 TTCTGGGGGC CCGCCCAGAT CAGGGCTGTG CTGGGAGCTG TGCCCCGTGG CCGCCTCCTG   7860  
               
               
                   
               
               
                 GTTCTGGACC TGTTTGCTGA GAGCCAGCCT GTGTATACCC GCACTGCCTC CTTCCAGGGC   7920  
               
               
                   
               
               
                 CAGCCCTTCA TCTGGTGCAT GCTGCACAAC TTTGGGGGAA ACCATGGTCT TTTTGGAGCC   7980  
               
               
                   
               
               
                 CTAGAGGCTG TGAACGGAGG CCCAGAAGCT GCCCGCCTCT TCCCCAACTC CACCATGGTA   8040  
               
               
                   
               
               
                 GGCACGGGCA TGGCCCCCGA GGGCATCAGC CAGAACGAAG TGGTCTATTC CCTCATGGCT   8100  
               
               
                   
               
               
                 GAGCTGGGCT GGCGAAAGGA CCCAGTGCCA GATTTGGCAG CCTGGGTGAC CAGCTTTGCC   8160  
               
               
                   
               
               
                 GCCCGGCGGT ATGGGGTCTC CCACCCGGAC GCAGGGGCAG CGTGGAGGCT ACTGCTCCGG   8220  
               
               
                   
               
               
                 AGTGTGTACA ACTGCTCCGG GGAGGCCTGC AGGGGCCACA ATCGTAGCCC GCTGGTCAGG   8280  
               
               
                   
               
               
                 CGGCCGTCCC TACAGATGAA TACCAGCATC TGGTACAACC GATCTGATGT GTTTGAGGCC   8340  
               
               
                   
               
               
                 TGGCGGCTGC TGCTCACATC TGCTCCCTCC CTGGCCACCA GCCCCGCCTT CCGCTACGAC   8400  
               
               
                   
               
               
                 CTGCTGGACC TCACTCGGCA GGCAGTGCAG GAGCTGGTCA GCTTGTACTA TGAGGAGGCA   8460  
               
               
                   
               
               
                 AGAAGCGCCT ACCTGAGCAA GGAGCTGGCC TCCCTGTTGA GGGCTGGAGG CGTCCTGGCC   8520  
               
               
                   
               
               
                 TATGAGCTGC TGCCGGCACT GGACGAGGTG CTGGCTAGTG ACAGCCGCTT CTTGCTGGGC   8580  
               
               
                   
               
               
                 AGCTGGCTAG AGCAGGCCCG AGCAGCGGCA GTCAGTGAGG CCGAGGCCGA TTTCTACGAG   8640  
               
               
                   
               
               
                 CAGAACAGCC GCTACCAGCT GACCTTGTGG GGGCCAGAAG GCAACATCCT GGACTATGCC   8700  
               
               
                   
               
               
                 AACAAGCAGC TGGCGGGGTT GGTGGCCAAC TACTACACCC CTCGCTGGCG GCTTTTCCTG   8760  
               
               
                   
               
               
                 GAGGCGCTGG TTGACAGTGT GGCCCAGGGC ATCCCTTTCC AACAGCACCA GTTTGACAAA   8820  
               
               
                   
               
               
                 AATGTCTTCC AACTGGAGCA GGCCTTCGTT CTCAGCAAGC AGAGGTACCC CAGCCAGCCG   8880  
               
               
                   
               
               
                 CGAGGAGACA CTGTGGACCT GGCCAAGAAG ATCTTCCTCA AATATTACCC CGGCTGGGTG   8940  
               
               
                   
               
               
                 GCCGGCTCTT GGTGATAGAT TCGCCACCAC TGGGCCTTGT TTTCCGCTAA TTCCAGGGCA   9000  
               
               
                   
               
               
                 GATTCCAGGG CCCAGAGCTG GACAGACATC ACAGGATAAC CCAGGCCTGG GAGGAGGCCC   9060  
               
               
                   
               
               
                 CACGGCCTGC TGGTGGGGTC TGACCTGGGG GGATTGGAGG GAAATGACCT GCCCTCCACC   9120  
               
               
                   
               
               
                 ACCACCCAAA GTGTGGGATT AAAGTACTGT TTTCTTTCCA CTTAAACTGA TGAGTCCCCT   9180  
               
               
                   
               
               
                 GGGTCTGTCA AAATGAGAAG GTCACTGCTG CCACGCTTGG GAGGACTCAG GGCTATAGCA   9240  
               
               
                   
               
               
                 TGGCCCTGGG GTGGGACCTG TTCTCCCATC CCTTGCCTCA CGTCCCTGTT TTTGTTTGTT   9300  
               
               
                   
               
               
                 TGTTTGTTTG TGACGGAGCC TTGGTCTGTT GCCCAGGCTT GAGTACAATG GCACAGTCTC   9360  
               
               
                   
               
               
                 GGCTCACTGC AACCTCCGCC TCCTGGGTTC AAGCAATTCT TGTGCCTCAG CCTCCCCGGT   9420  
               
               
                   
               
               
                 AGCTGGGACT ATAGGCATGC ACCACCACAC CAGGCTAATT TTTTTTTTTC CAAGATGGAG   9480  
               
               
                   
               
               
                 TCTTGCTCTG TCGCCCAGGT TGGAGTTTAG TGGCACCATA TTGGTTTACT GCAACCTCTG   9540  
               
               
                   
               
               
                 CCTCCCGGGT TCAAGCAATT CTCCTGCCTC AGTCTACCAG GGAGTTAGGA CTACGGGCCT   9600  
               
               
                   
               
               
                 GTGCCATCAC GCCTGGCTAA TTTTTGTATT TTTCATAGAG ATAAGGTTTC ACCATGTTGG   9660  
               
               
                   
               
               
                 CCAGGCTGGT CTTTAACTCC TGAACTCAAG TGATCCACCT GCCTCGGCCT TCCAAAGTGC   9720  
               
               
                   
               
               
                 TGGGATTACA GGAGTGAGCC ACCGTGCCCG GCCATGTCTC TCTTTTTAAC ACTAATGTTA   9780  
               
               
                   
               
               
                 CCCTGACCTT TGAACGTAGA ATGCCCTTCT GTTGCAGGAA AACCTCTTTT CAAACCATGT   9840  
               
               
                   
               
               
                 TTGTCCTTTG CTGGCATGCC ACAGCAACAG TCACCAACAC AGAAGACTTC TGTGACCAAA   9900  
               
               
                   
               
               
                 TATTTGGAGG ATTTTCCCCA CACACACCAA GCAGCAGACA TCAGCTGGGT GTCCTCCAAT   9960  
               
               
                   
               
               
                 TCAGTTCCAA TGTAATCAAC CAGAGACAGC ATCAGATCCC ACAGGGTTAG GGTGCAGATC  10020  
               
               
                   
               
               
                 CATGAGACCA CCCCCTCCTT CCCAACGGTT ACAAGTCCTG ATCCCTGGAA CTTCTGACTA  10080  
               
               
                   
               
               
                 ACTGGCTTCA AGTTGGAGTT CCCATGACCC CCTTCCCCTC TTTGGAGTCA ACTCATTTGC  10140  
               
               
                   
               
               
                 GACAGTGACC CACGAAACAC AGGGAAACCC TTATTATGTT TATTGCTTTA TTACAGAGGA  10200  
               
               
                   
               
               
                 AAAAAATTTT TTTCTTTCTT TTTTGAGACA GGGTCTCACT CTGTCATCCA GAATGACTGC  10260  
               
               
                   
               
               
                 AGTGGCAGGA TCTGGCTCCG TCACCCAGGC TGGAGTGCAG TGGCATGATC TCGGCTCACT  10320  
               
               
                   
               
               
                 ACAGCCTCCA TCCCCCCCAA ACCCCACGCC TCAGCGCCCC ACCCCGCAAG TGGCTGGGAC  10380  
               
               
                   
               
               
                   
               
               
                 (2) INFORMATION FOR SEQ ID NO: 4:  
               
               
                   
               
               
                      (i) SEQUENCE CHARACTERISTICS:  
               
               
                           (A) LENGTH: 20 amino acids  
               
               
                           (B) TYPE: amino acid  
               
               
                           (D) TOPOLOGY: linear  
               
               
                   
               
               
                     (ii) MOLECULE TYPE: peptide  
               
               
                   
               
               
                    (iii) HYPOTHETICAL: NO  
               
               
                   
               
               
                      (v) FRAGMENT TYPE: N-terminal  
               
               
                   
               
               
                     (vi) ORIGINAL SOURCE:  
               
               
                           (A) ORGANISM: Homo sapiens  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Modified-site  
               
               
                           (B) LOCATION: 10  
               
               
                   
               
               
                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:  
               
               
                   
               
               
                 Asp Glu Ala Arg Glu Ala Ala Ala Val Arg Ala Leu Val Ala Arg  
               
               
                  1               5                   10                  15  
               
               
                   
               
               
                 Leu Leu Gly Pro Gly  
               
               
                                  20  
               
               
                   
               
               
                   
               
               
                 (2) INFORMATION FOR SEQ ID NO: 5:  
               
               
                   
               
               
                      (i) SEQUENCE CHARACTERISTICS:  
               
               
                           (A) LENGTH: 18 amino acids  
               
               
                           (B) TYPE: amino acid  
               
               
                           (D) TOPOLOGY: linear  
               
               
                   
               
               
                     (ii) MOLECULE TYPE: peptide  
               
               
                   
               
               
                    (iii) HYPOTHETICAL: NO  
               
               
                   
               
               
                      (v) FRAGMENT TYPE: N-terminal  
               
               
                   
               
               
                     (vi) ORIGINAL SOURCE:  
               
               
                           (A) ORGANISM: Homo sapiens  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Modified-site, glycosylated or phosphorylated,  
               
               
                                wherein Xaa may be any amino acid residue, preferably  
               
               
                                Arg.  
               
               
                           (B) LOCATION: 16  
               
               
                   
               
               
                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5:  
               
               
                   
               
               
                 Lys Pro Gly Leu Asp Thr Tyr Ser Leu Gly Gly Gly Gly Ala Ala Xaa Val  
               
               
                  1               5                   10                  15  
               
               
                   
               
               
                 Arg  
               
               
                   
               
               
                   
               
               
                 (2) INFORMATION FOR SEQ ID NO: 6:  
               
               
                   
               
               
                      (i) SEQUENCE CHARACTERISTICS:  
               
               
                           (A) LENGTH: 15 amino acids  
               
               
                           (B) TYPE: amino acid  
               
               
                           (D) TOPOLOGY: linear  
               
               
                   
               
               
                     (ii) MOLECULE TYPE: peptide  
               
               
                   
               
               
                    (iii) HYPOTHETICAL: NO  
               
               
                   
               
               
                      (v) FRAGMENT TYPE: internal  
               
               
                   
               
               
                     (vi) ORIGINAL SOURCE:  
               
               
                           (A) ORGANISM: Homo sapiens  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Modified-site, glycosylated or phosphorylated,  
               
               
                                wherein Xaa may be any amino acid residue, preferably Ala  
               
               
                           (B) LOCATION: 12  
               
               
                   
               
               
                     (ix) FEATURE:  
               
               
                           (A) NAME/KEY: Modified-site, glycosylated or phosphorylated,  
               
               
                                wherein Xaa may be any amino acid residue, preferably Ser  
               
               
                           (B) LOCATION: 14  
               
               
                   
               
               
                     (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6:  
               
               
                   
               
               
                 Trp Arg Leu Leu Leu Thr Ser Ala Pro Ser Leu Xaa Thr Xaa Pro  
               
               
                 1               5                   10                  15