CHITOTRIOSIDASE

Abstract: Nucleic acid sequences for chitotriosidase are provided. Methods of detecting altered expression of tissue remodeling proteins and diagnosing tissue remodeling disorders are also provided.

Description: CHITOTRIOSIDASE BACKGROUND OF THE INVENTION The generation or destruction of tissue requires constant reorganization and restructuring of the extracellular matrix (ECM) components including interstitial collagens, basement membrane collagen, fibronectin, lamimn, aggrecan, and vaπous proteoglycans. Heinegard and Oldberg, FASEB J. 1989, 3, 2042-2051; Woessner FASEB J 1991, 5, 214-2154. Normal types of remodeling processes include embryonic development, post-partum involution of the uterus, ovulation, wound healing, and bone and growth plate remodeling. Woessner et al. Steroids 1989, 54, 49M99; Weeks et al. Biochim Biophys Acta 1976, 445, 205-214; Lepage and Gache EMBO J. 1990, 9, 3003-3012; Wride and Sanders Dev-Dyn. 1993. 198(3) 225-39. Similar processes also occur in disease states such as joint destruction in rheumatoid and osteoarthritis, peπodontia and tumor cell metastasis. Thompson and Oegema J Bone Joint Stirg. 1979. 61, 407-16; Reynolds et al. Adv-Dent-Res. 1994, 8(2) 312-9. One example of these processes is the migration of macrophages to the site of inflammation as in the case of synovial tissue in rheumatoid arthntis Cutolo et al. Clin. and Exper. Rheum. 1993, 11, 331-339. The ECM components are regulated, in both normal and disease states, by various exogenous and endogenous factors. For example, in tumor formation, the differentiation state of the cell can increase the rate of degradation of the ECM. Benya Pathol. Immunopathol. Res. 1988, 7, 51-54 Likewise, the presence of metalloproteinases or their inhibitors can alter the composition of the ECM. An imbalance of metalloproteinases and tissue inhibitors of matrix metalloproteinases (TIMP) has been shown to contπbute to the pathogenesis of osteoarthritis. Dean et al J. Clin. Invest. 1989, 84: 678-685 Cytokines, growth factors, and the extracellular environment can all contπbute to the alteration of the ECM. Tyler Biochem J. 1985, 227, 869-878; Dinarello Sem Immunol. 1992, 4, 133-145; McConnell et al.7. Cell Biol. 1987, 105, 1087-98. The growth of cartilage and bone is actualized by cells such as articular chondrocytes and osteoblasts. The main function of these cells in immature tissue is the deposition and remodeling of the cartilage or bone matπx. In adult tissue, these cells maintain this matrix in order to ensure its proper function. In both cases, this encompasses secretion of the extracellular components as well as secretion of proteins involved in the turnover of the ECM. A major species of protein secreted by these cells and involved in the turnover of the ECM are the metalloproteinases. Woessner FASEB J. 1991, 5, 214-2154. A new type of secretory glycoprotein has also been identified in human cartilage, osteoblasts. synovial cells, sheep and bovine oviduct and mammary cells, and macrophages. Nyrikos and Golds Biochem J. 1990, 268, 265-268; Hakala et al. J. Biol. Chem. 1993, 268(34) 25803-25810; Johansen et al. J. Bone and Min. Res. 1992, 7(5) 501-51 1; Rejman and Hurley Biochem. Biophys. Res. Commun. 1988. 150, 329-334; DeSouza and Murray Endocrinology 1995, 136(6) 2485-2496; Hollak et al. J. Clin. Invest. 1994, 93, 1288-92; Arias et al. Biol. of Reproduction 1994, 51, 685-694. Chitotriosidase shares regions of significant homology to the bacterial and fungal chitinases. Chitinases are enzymes that hydrolyze glycosidic bonds. They bear a subtle similarity to lysozymes from mammals and function as endoglycosidases with a specificity for N-acetyl-glucosamine linkages. However, these types of chitin-like structures, homopolymers of N-acetyl-glucosamine. are not normally encountered in mammalian tissue. The human cartilage glycoprotein, HC gp-39, is a protein with an apparent molecular weight of approximately 39 kDa secreted by both articular chondrocytes and synovial fibroblasts. Nyrikos and Golds Biochem J. 1990, 268, 265-268; Hakala et al. J. Biol. Chem. 1993, 268(34), 25803-25810. This protein has been described as a marker for joint injury, appearing in the blood and synovial fluid from patients diagnosed with rheumatoid arthritis. Johansen et al. British J. of Rheumatology 1993, 32, 949-955. The gene encoding this protein has been cloned and is expressed specifically in cartilage and synovial cells of rheumatic joints. Hakala et al. J. Biol. Chem. 1993, 268(34), 25803-25810. The protein YKL-40 has also been identified as one of the major secretory products of cultured human osteoblastic cells (osteocarcinoma cell line MG-63) expressed in response to 1 ,25-dihydroxyvitamin D3 stimulation. Johansen et al. J. Bone and Min. Res. 1992, 7(5), 501-51 1; Johansen et al. Br. J. Rheumatol. 1993, 32, 949-55. The N-terminal portion of YKL-40 was sequenced and found to be identical to HC gp-39. Upon further sequencing, YKL-40 and HC gp-39 were found to be identical. Chitotriosidase is an enzyme which has been identified as a member of this family homologous to the chitinases. Renkema et al. J. Biol Chem. 1995, 27C, 2198-2202; Hollak et al. J. Clin. Invest. 1994, 93, 1288-92. This protein also has an apparent molecular weight of 39 kDa and shares N-terminal homologies with HC gp-39, the bovine mammary protein, and several bacterial chitinases. Activity of this enzyme was originally detected from cells of patients afflicted with Gaucher Disease (GD). Gaucher Disease is an inherited deficiency in the activity of glucocerebrosidase, a lysosomal hydrolase. This defect results in an accumulation of glucosylceramide (glucocerebroside) in the lysosomes of macrophages. Accumulation of lipid-laden macrophages results in hepatosplenomegaly, bone lesions, and neurological anomalies. After morphological differentiation of monocytes into macrophages in culture, the cells begin to produce and secrete increasing amounts of chitotriosidase. This increase is, on average, 600 times greater in GD patients than in patients with other pathological conditions. The elevation in chitotriosidase activity can be effectively reduced, however, upon initiation of enzyme supplementation therapy. Unlike the other member of the family of proteins homologous to chitinase, chitotriosidase has chitolytic activity. Like the bacterial enzyme, it has the ability to degrade chitin azure, a polymer of beta- 1 -4-linked N-acetylglucosamine moieties. The full length cDNA of chitotriosidase has now been found. This protein, along with HC gp-39, is believed to be involved in tissue remodeling in the mammalian cell and thus serve as useful tools in the development of therapeutics and diagnostics for tissue remodeling disorders. Summary ofthe Invention In accordance with one aspect of the present invention, there is provided a mature polypeptide which is chitotriosidase, as well as biologically active and diagnostically or therapeutically useful fragments, analogs and derivatives thereof. The polypeptide of the present invention is of human origin. In accordance with another aspect of the present invention, there are provided isolated nucleic acid molecules encoding human chitotriosidase, including mRNAs, DNAs, cDNAs, genomic DNAs as well as analogs and biologically active and diagnostically or therapeutically useful fragments thereof. In accordance with yet a further aspect of the present invention, there is provided a process for producing chitotriosidase by recombinant techniques comprising culturing recombinant prokaryotic and or eukaryotic host cells, containing a human nucleic acid sequence of chitotriosidase, under conditions promoting expression of these proteins and subsequent recovery of these proteins. In accordance with yet a further aspect of the present invention, there is provided a process for utilizing chitotriosidase polypeptide, for therapeutic purposes, for example, in the treatment of tissue remodeling disorders. In accordance with yet a further aspect of the present invention, there is provided a process for utilizing chitotriosidase polypeptide, for diagnostic purposes, for example, in the diagnosis of tissue remodeling disorders. These and other aspects of the present invention should be apparent to those skilled in the art from the teachings herein. Brief Description of the Drawings Figure 1 provides the coding sequence (SEQ ID NO: 1) and deduced amino acid sequence (SEQ ID NO: 2) of the polypeptide chitotriosidase. Figure 2 provides the coding sequence (SEQ ID NO: 3) and deduced amino acid sequence (SEQ ID NO: 4) of the polypeptide chitotriosidase. Detailed Description ofthe Invention In accordance with one aspect of the present invention, there is provided an isolated nucleic acid sequence which encodes for the chitotriosidase (SEQ ID NO: 1 ). The deduced amino acid sequence of this protein (SEQ ID NO: 2) is shown in Figure 1. Further, the full length cDNA of the clone encoding chitotriosidase was deposited as ATCC Deposit No. 69953 on November 29, 1995. Sequencing of this full length clone encoding chitotriosidase revealed a nucleotide sequence that is 54% homologous to HC gp-39. This gene contains an open reading frame coding for a 395 amino acid peptide that is 52% identical (68% amino acid similarity) to HC gp-39. In accordance with another aspect of the invention the following nucleotide sequence variations of the chitotriosidase in Figure 1 (SEQ ID NO: 1) are provided, singly or preferably in combination (nucleotide numbers correspond to Figure 2 (SEQ ID NO: 1): nucleotide 330 is a G, or an insertion of 6 nucleotides, ggtaca, or, nucleotide 1253 is a C, or nucleotide 1426 is deleted (C deleted), or nucleotide 1442 is deleted (C deleted), or nucleotide 1495 is deleted (C deleted), or nucleotide 1632 is a G, or nucleotide 1634 is a G. Skilled artisans will readily be able to determine the concomitant amino acid differences caused by these nucleotide changes from Figure 2 (SEQ ED NO: 1) or whether they are silent changes using methods routine in the art. Further provided is a preferred sequence variant depicted in Figure 2 (SEQ ID NO: 3 and 4). The nucleic acid sequences of the present invention may be in the form of RNA or in the form of DNA, which DNA includes cDNA, genomic DNA, and synthetic DNA. The DNA may be double-stranded or single-stranded, and if single stranded may be the coding strand or non- coding (anti-sense) strand. The coding sequence which encodes the chitotriosidase protein may be identical to the coding sequence shown in Figure 1 or that of the deposited clone or may be a different coding sequence which coding sequence, as a result of the redundancy or degeneracy of the genetic code, encodes the same protein as the DNA of Figure 1 or the deposited cDNAs. The present invention includes polynucleotides encoding the same mature polypeptide as shown in Figure 1 or the same mature polypeptide encoded by the cDNA of the deposited clone Further, the inventions includes vaπants of such polynucleotides that encode a fragment, deπvative or analog of the polypeptides of Figure 1 or the polypeptide encoded by the cDNA of the deposited clone Among vaπants in this regard are vaπants that differ from the aforementioned polynucleotides by nucleotide substitutions, deletions or additions The substitutions, deletions or additions may inolve one or more nucleotides The vaπants may be altered in coding or non-coding regions or both Alterations in the coding regions may produce conservative amino acid sustitutions, deletions or additions Vaπants of the invention may have a sequence that occurs in nature or they may have a sequence that does not occur naturally As herein above indicated, the polynucleotide may have a coding sequence which is a naturally occurring allelic vanant of the coding sequences shown in Figure 1 or of the coding sequences of the deposited clone As known in the art, an allelic vanant is an alternate form of a polynucleotide sequences which may have a substitution, deletion or addition of one or more nucleotides Among the particularly preferred embodiments of the invention in this regard are polynucleotides encoding polypeptides having the amino acid sequences of chitotπosidase set out in Figure 1 or the amino acid sequence of chitotπosidase of the cDNA of the deposited clone, vanants, analogs, denvatives and fragments thereof, and fragments of the vaπants, analogs and deπvatives Further particularly preferred in this regard are polynucleotides encoding chitotπosidase vaπants. analogs, deπvatives and fragments, and vaπants, analogs and deπvatives of the fragments, which have the ammo acid sequence of the chitotπosidase polypeptide of Figure 1 or of the deposit in which several, a few, 5 to 10, 1 to 5, 1 to 3, 2, 1 or no amino acid residues are substituted, deleted or added, in any combination Especially preferred among these are silent substitutions, additions and deleUons, which do not alter the properties and activities of the chitotnosidase Also especially preferred in this regard are conservative substitutions most highly preferred are polypeptides having the amino acid sequence of Figure 1 or of the deposit, without substitutions Further preferred embodiments of the invention are polynucleotides that are more than 85% identical to a polynucleotide encoding the chitotπosidase polypeptide having the amino acid sequence set out in Figure 1, or vaπants, close homologs, deπvatives and analogs thereof, as descπbed above Alternatively, most highly preferred are polynucleotides that compπse a region that is more than 85% identical to a polynucleotide encoding the chitotπosidase polypeptide of the cDNA of the deposited clone In this regard, polynucleotides more than 90% identical to the same are particularly preferred, and among these particularly preferred polynucleotides, those with 95% or more identity are especially preferred Furthermore, those with 97% or more identity are highly preferred among those with 95% or more identity, and among these those with 98% or more and 99% or more identity are particularly highly prefered, with 99% or more being the more prefeπed Also particularly preferred in this regard are polynucleotides encoding a polypeptide having the amino acid sequence of the chitotriosidase set out in Figure 1 or of the deposited clone As set out elsewhere herein, the polynucleotide may encode the polypeptide in a continuous region or in a plurality of two or more discontinuous exons, and it may compnse additional regions as well, which are unrelated to the coding region or regions Most highly preferred in this regard are polynucleotides that compnse a region that is more than 85% identical to the chitotπosidase-encoding portion of the pol nucelotide set out in Figure 1 Alternatively, most highly preferred are polynucleotides that compπse a region that is more than 85% identical to the chitotπosidase-encoding portion of the cDNA of the deposited clone Among such polynucleotides, those more than 90% identical to the same are particularly preferred, and, among these particularly preferred polynucleotides, those with 95% or more lentity are especially preferred Furthermore, those with 97% or more identity are highly preferred among those with 95% or more identity, and among these those with 98% or more and 99% or more identity are particularly highly preferred, with 99% or more being the more preferred of these The present invention also includes polynucleotides in which the sequence encoding the mature polypeptide is fused in the same reading frame to additional sequences Such sequences include signal sequences, which facilitate transport of the nascent protein into the endoplasmic reticulum, pro-sequences that are associated with inactive precursor forms of the polypeptide, which may facihtate trafficking of the protein in a cell or out of a cell or may unprove persistence of the protein in a cell or in an extracellular compartment. Such sequences also may be added to facihtate production and purification, or to add additional functional domains, as discussed elsewhere herein The deposιt(s) referred to herein will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Micro-organisms for purposes of Patent Procedure These deposits are provided merely as convenience to those of skill in the art and are not an admission that a deposit is required under 35 U S C §112. The sequence of the polynucleotides contained in the deposited matenals, as well as the amino acid sequence of the polypeptides encoded thereby, are incorporated herein by reference and are controlling in the event of any conflict with any descπption of sequences herein A license may be required to make, use or sell the deposited matenals, and no such license is hereby granted The present invention further relates to the chitotπosidase polypeptide which has the deduced amino acid sequence (SEQ ID NO 2) of Figure 1 or which has the amino acid sequence encoded by the deposited cDNA. as well as fragments, analogs and deπvatives of such polypeptide The teπns "fragment," "deπvative" and "analog" when referring to the polypeptide of Figure 1 or that encoded by the deposited cDNA, means a polypeptide which retains essentially the same biological function or activity as such polypepnde Thus, an analog includes a proprotein which can be activated by cleavage of the proprotein portion to produce an active mature polypeptide The polypeptide of the present invention may be a recombinant polypeptide. a natural polypeptide or a synthetic polypeptide, preferably a recombinant polypeptide The fragment, denvative or analog of the polypeptide of Figure 1 or that encoded by the deposited cDNA may be (0 one in which one or more ot the ammo acid residues are substituted with a conserved or non-conserved ammo acid residue (preferably a conserved ammo acid residue) and such substituted ammo acid residue may or may not be one encoded by the genetic code, or (u) one in which one or more of the ammo acid residues mcludes a substituent group, or Oil) one in which the mature polypeptide is fused with another compound, such as a compound to increase the half-life of the polypeptide (for example, polyethylene glycol), or (iv) one in which the additional ammo acids are fused to the mature polypeptide, such as a leader or secretory sequence or a sequence which is employed for purification of the mature polypeptide or a proprotein sequence Such fragments, denvatives and analogs are deemed to be withm the scope of those skilled in the art from the teachings herem The polypeptide and nucleic acid sequences of the present mvention are preferably provided m an isolated form, and preferably are purified to homogeneity The term "isolated" means that the matenal is removed from its onginal environment (e g , the natural environment if it is naturally occurring) For example, a naturally-occurπng nucleic acid sequence or polypeptide present m a living animal is not isolated, but the same nucleic acid sequence or polypeptide, separated from some or all of the coexistmg matenals m the natural system, is isolated Such nucleic acid sequences could be part ofa vector and/or such nucleic acid sequences or polypeptides could be part of a composition, and still be isolated in that such vector or composition is not part of its natural environment The present invention also relates to vectors which include one of the nucleic acid sequences of the present invention, host cells which are genetically engineered with vectors of the invention and the production of polypeptides of the invention by recombinant techniques. Host cells are genetically engineered (transduced or transformed or transfected) with the vectors of this invention which may be, for example, a cloning vector or an expression vector. The vector may be, for example, in the form of a plasmid. a viral particle, a phage, etc. The engineered host cells can be cultured in conventional nutrient media modified as appropriate for activating promoters, selecting transformants or amplifying the chitotriosidase genes. The culture conditions, such as temperature, pH and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan. The nucleic acid sequences of the present invention may be employed for producing polypeptides by recombinant techniques. The nucleic acid sequences may be included in any one of a variety of expression vectors for expressing a polypeptide. Such vectors include chromosomal, nonchromosomal and synthetic DNA sequences, e.g., derivatives of SV40; bacterial piasmids; phage DNA; baculovirus; yeast piasmids; vectors derived from combinations of piasmids and phage DNA, viral DNA such as vaccinia, adenovirus, fowl pox virus, and pseudorabies. However, any other vector may be used as long as it is replicable and viable in the host. The DNA sequence of chitotriosidase can be inserted into the vector by a variety of procedures. In general, the DNA sequence is inserted into an appropriate restriction endonuclease site(s) by procedures known in the art. Such procedures and others are deemed to be within the scope of those skilled in the art. The DNA sequence in the expression vector is operatively linked to an appropriate expression control sequence(s) (promoter) to direct mRNA synthesis. As representative examples of such promoters, there may be mentioned: LTR or SV40 promoter, the E. coli lac or trp, the phage lambda P promoter and other promoters known to control expression of genes in prokaryotic or eukaryotic cells or their viruses. The expression vector also contains a ribosome binding site for translation initiation and a transcription terminator. The vector may also include appropriate sequences for amplifying expression. In addition, the expression vectors preferably contain one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells such as dihydrofolate reductase or neomycin resistance for eukaryotic cell culture, or such as tetracycline or ampicillin resistance in E. coli. The vector contammg the appropπate DNA sequence as hereinabove descπbed, as well as an appropπate promoter or control sequence, may be employed to transform an appropπate host to permit the host to express the protein As representative examples of appropπate hosts, there may be mentioned bacteπal cells, such as E colt, Streptomyces, Salmonella typhimunum fungal cells, such as yeast, insect cells such as Drosophila S2 and Spodoptera Sf9, animal cells such as CHO, COS or Bowes melanoma, adenoviruses, plant cells, etc The selection of an appropπate host is deemed to be withm the scope of those skilled in the art from the teachings herem More particularly, the present mvention also mcludes recombmant constructs compnsmg one or more of the sequences as descπbed above The constructs compπse a vector, such as a plasmid or viral vector, mto which a sequence encodmg chitotπosidase has been inserted, a forward or reverse oπentation In a preferred aspect of this embodiment, the construct further compnses regulatory sequences, mcludmg, for example, a promoter, operably linked to the sequence Large numbers of suitable vectors and promoters are known to those of skill in the art, and are commercially available The following vectors are provided by way of example, Bacteπal pQE70, pQE60, pQE-9 (Qiagen), pBS, pDIO, phagescπpt, psιX174, pbluescnpt SK, pbsks, pNH8A, pNHlόa, pNH18A, pNH46A (Stratagene), ptrc99a, pKK223-3, pKK233-3, pDR540, pRJT5 (Pharmacia), Eukaryotic pWLNEO, pSV2CAT, pOG44, pXTl, pSG (Stratagene) pSVK3, pBPV, pMSG, pSVL (Pharmacia) However, any other plasmid or vector may be used as long as they are replicable and viable in the host Promoter regions can be selected from any des ed gene usmg CAT (chloramphenicol transferase) vectors or other vectors with selectable markers Two appropriate vectors are pKK232-8 and pCM7 Particular named bacteπal promoters mclude lad, lacZ, T3, T7, gpt, lambda P R , P and trp Eukaryotic promoters include CMV immediate early, HSV thymidine kmase, early and late SV40, LTRs from retrovirus, and mouse metallothιoneιn-I Selection of the appropπate vector and promoter is well with the level of ordinary skill the art In a further embodiment, the present mvention relates to host cells contammg the above- descπbed constructs The host cell can be a higher eukaryotic cell, such as a mammalian cell, or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacteπal cell. Introduction of the construct mto the host cell can be effected by calcium phosphate transfecuon, DEAE-Dextran mediated transfection, or electroporation (Davis, L , Dibner, M , Battey, I , Basic Methods in Molecular Biology, (1986)) The constructs in host cells can be used in a conventional manner to produce the gene product encoded by the recombinant sequence. Alternatively, polypeptides can be synthetically produced by conventional peptide synthesizers. Mature proteins can be expressed in mammalian cells, yeast, bacteria, or other cells under the control of appropriate promoters. Cell-free translation systems can also be employed to produce such proteins using RNAs derived from the DNA constructs of the present invention. Appropriate cloning and expression vectors for use with prokaryotic and eukaryotic hosts are described by Sambrook et al.. Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor, N.Y., (1989), the disclosure of which is hereby incoφorated by reference. Transcription of the DNA encoding a protein by higher eukaryotes is increased by inserting an enhancer sequence into the vector. Enhancers are cis-acting elements of DNA, usually about from 10 to 300 bp that act on a promoter to increase its transcription. Examples including the SV40 enhancer on the late side of the replication origin bp 100 to 270, a cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers. Generally, recombinant expression vectors will include origins of replication and selectable markers permitting transformation of the host cell, e.g., the ampicillin resistance gene of E. coli and S. cerevisiae TRP1 gene, and a promoter derived from a highly-expressed gene to direct transcription of a downstream structural sequence. Such promoters can be derived from operons encoding glycolytic enzymes such as 3-phosphoglycerate kinase 0?GK), a-factor, acid phosphatase, or heat shock proteins, among others. The heterologous structural sequence is assembled in appropriate phase with translation initiation and termination sequences, and preferably, a leader sequence capable of directing secretion of translated protein into the periplasmic space or extracellular medium. Optionally, the heterologous sequence can encode a fusion protein including an N-terminal identification peptide imparting desired characteristics, e.g., stabilization or simplified purification of expressed recombinant product. Useful expression vectors for bacterial use are constructed by inserting a structural DNA sequence encoding a desired protein together with suitable translation initiation and termination signals in operable reading phase with a functional promoter. The vector will comprise one or more phenotypic selectable markers and an origin of replication to ensure maintenance of the vector and to, if desirable, provide amplification within the host. Suitable prokaryotic hosts for transformation include E. coli. Bacillus subtilis. Salmonella typhimurium and various species withm the genera Pseudomonas, Streptomvces, and Staphylococcus. although others may also be employed as a matter of choice As a representative but nonlimitmg example, useful expression vectors for bacteπal use can compπse a selectable marker and bactenal ongin of replication deπved from commercially available piasmids compnsmg genetic elements of the well known clonmg vector pBR322 (ATCC 37017) Such commercial vectors mclude, for example, pKK223-3 (Pharmacia Fme Chemicals, Uppsala, Sweden) and GEM1 (Promega Biotec, Madison, WI, USA) These pBR322 "backbone" sections are combmed with an appropπate promoter and the structural sequence to be expressed Following transformation of a suitable host strain and growth of the host stra to an appropπate cell density, the selected promoter is induced by appropπate means (e g , temperature shift or chemical duction) and cells are cultured for an additional penod Cells are typically harvested by centπftigation, disrupted by physical or chemical means, and the resultmg crude extract retained for further purification Microbial cells employed in expression of protems can be disrupted by any convenient method, mcludmg freeze-thaw cychng, sonication, mechanical disruption, or use of cell lys g agents, such methods are well know to those skilled in the art Vaπous mammalian cell culture systems can also be employed to express recombinant protems Examples of mammahan expression systems mclude the COS-7 lmes of monkey kidney fibroblasts, descπbed by Gluzman Cell 1981, 23, 175, and other cell hnes capable of expressmg a compatible vector, for example, the C127, 3T3, CHO, HeLa and BHK cell hnes Mammalian expression vectors will compπse an ongin of replication, a suitable promoter and enhancer, and also any necessary πbosome bmdmg sites, polyadenylation site, splice donor and acceptor sites, transcπptional termination sequences, and 5' flanking nontranscnbed sequences DNA sequences deπved from the SV40 splice, and polyadenylation sites may be used to provide the required nontranscnbed genetic elements Polypeptides can then be recovered and punfied from recombmant cell cultures by methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic mteraction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography Protem refoldmg steps can be used, as necessary, in completmg configuration of the mature protem Finally, high performance liquid chromatography (HPLC) can be employed for final purification steps It is now believed that chitotriosidase is involved in tissue remodeling. Human chitotriosidase is homologous to microbial chitinases. In particular the protein shares a similar cysteine motif and homology to the active site of microbial chitinases. Accordingly, chitotriosidase can be used in the development of treatments for tissue remodeling diseases and in the diagnosis of these diseases. It is believed that human serum chitotriosidase is an extracellular matrix remodeling factor involved in the remodeling process occurring in diseases. Activity against chitotriosidase substrates is indicative of a function in degrading extracellular matrix substrates with a similar carbohydrate structure to chitin. Chitotriosidase was found during morphological differentiation of monocytes into macrophages. These macrophages can be associated with different sites of inflammation that occur in various disease states, including but not limited to atheroschlerosis, rheumatoid arthritis, acute inflammatory disease, and inflammation due to injury. Accordingly, it is believed that human serum chitotriosidase is a useful target for the inhibition of tissue remodeling disease processes. Purified human serum chitotriosidase protein could thus be tested for various in vitro and in vivo assays for tissue remodeling to determine the extent to which this protein promotes these processes. Therapeutics would be defined as molecules that specifically inhibit chitotriosidase function as it relates to tissue remodeling activity in these assays and includes neutralizing antibodies. Chitotriosidase can also be used in the production of antibodies. The proteins, their fragments or other derivatives, or analogs thereof, or cells expressing them can be used as an immunogen to produce antibodies thereto. These antibodies can be, for example, polyclonal or monoclonal antibodies. The present invention also includes chimeric, single chain, and humanized antibodies, as well as Fab fragments, or the product of an Fab expression library. Various procedures known in the art may be used for the production of such antibodies and fragments. Antibodies generated against these proteins can be obtained by direct injection of the protein into an animal, preferably a nonhuman. The antibody so obtained will then bind the protein itself. In this manner, even a sequence encoding only a fragment of the protein can be used to generate antibodies binding the whole native protein. Such antibodies can then be used to isolate these proteins from tissue expressing these proteins. For preparation of monoclonal antibodies, any technique which provides antibodies produced by continuous cell line cultures can be used. Examples include the hybridoma technique (Kohler and Milstein Nature 1975, 256, 495-497), the trioma technique, the human B-cell hybridoma technique (Kozbor et al. Immunology Today 1983, 4, 72), and the EBV-hybridoma technique to produce human monoclonal antibodies (Cole et al. in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., 1985, pp. 77-96). Techniques described for the production of single chain antibodies (U.S. Patent 4,946,778) can be adapted to produce single chain antibodies to immunogenic chitotriosidase protein product. Also, transgenic mice may be used to express humanized antibodies to immunogenic chitotriosidase protein products. This invention is also related to the use of these chitotriosidase protein as a diagnostic. Detection of a mutated forms of chitotriosidase will allow a diagnosis of a tissue remodeling disease such as rheumatoid arthritis or atherosclerosis. Individuals caπying mutations in one or more of chitotriosidase protein may be detected at the DNA level by a variety of techniques. Nucleic acids for diagnosis may be obtained from a patient's cells, such as from blood, urine, saliva, tissue biopsy and autopsy material. The genomic DNA may be used directly for detection or may be amplified enzymatically by using PCR (Saiki et al.. Nature 1986, 324, 163-166) prior to analysis. RNA or cDNA may also be used for the same purpose. As an example, PCR primers complementary to the nucleic acid encoding chitotriosidase can be used to identify and analyze mutations in this protein. For example, deletions and insertions can be detected by a change in size of the amplified product in comparison to the normal genotype. Point mutations can be identified by hybridizing amplified DNA to radiolabeled RNA or altematively, radiolabeled antisense DNA sequences. Perfectly matched sequences can be distinguished from mismatched duplexes by RNase A digestion or by differences in melting temperatures. Sequence differences between the reference gene and genes having mutations may be revealed by the direct DNA sequencing method. In addition, cloned DNA segments may be employed as probes to detect specific DNA segments. The sensitivity of this method is greatly enhanced when combined with PCR. For example, a sequencing primer is used with double- stranded PCR product or a single-stranded template molecule generated by a modified PCR. The sequence determination is performed by conventional procedures with radiolabeled nucleotide or by automatic sequencing procedures with fluorescent-tags. Genetic testing based on DNA sequence differences may be achieved by detection of alteration in electrophoretic mobility of DNA fragments in gels with or without denaturing agents. Small sequence deletions and insertions can be visualized by high resolution gel electrophoresis. DNA fragments of different sequences may be distinguished on denaturing formamidine gradient gels in which the mobilities of different DNA fragments are retarded in the gel at different positions accordmg to their specific meltmg or partial meltmg temperatures (see, e g , Myers et dl Science 1985, 230, 1242) Sequence changes at specific locations may also be revealed by nuclease protection assays, such as RNase and S 1 protection or the chemical cleavage method (e g , Cotton et al PNAS USA 1985, 85, 4397-4401 Thus, the detection of a specific DNA sequence may be achieved by methods such as hybndization, RNase protection, chemical cleavage, direct DNA sequencmg or the use of restnction enzymes, (e g , Restnction Fragment Length Polymorphisms (RFLP)) and Southern blotting of genomic DNA In addition to more conventional gel-electrophoresis and DNA sequencmg, mutations can also be detected by in situ analysis The present invention also relates to a diagnostic assay for detecting altered levels of chitotπosidase protem m vaπous tissues Assays used to detect levels of these protems in a sample deπved from a host are well-known to those of skill in the art and mclude radioimmunoassays, competitive-binding assays. Western Blot analysis and preferably an ELISA assay An ELISA assay initially compnses preparing an antibody specific to chitotriosidase protein, preferably a monoclonal antibody In addition, a reporter antibody is prepared agamst the monoclonal antibody To the reporter antibody is attached a detectable reagent such as radioactivity, fluorescence or in this example a horseradish peroxidase enzyme A sample is removed from a host and mcubated on a solid support, e g , a polystyrene dish, that bmds the protems m the sample Any free protem bmdmg sites on the dish are then covered by mcubatmg with a non-specific protem like BSA Next, the monoclonal antibody is incubated in the dish dunng which time the monoclonal antibodies attach to chitotπosidase protein attached to the polystyrene dish All unbound monoclonal antibody is washed out with buffer The reporter antibody linked to horseradish peroxidase is now placed m the dish resultmg in bmdmg of the reporter antibody to any monoclonal antibody bound to chitotπosidase protem Unattached reporter antibody is then washed out. Peroxidase substrates are then added to the dish and the amount of color developed in a given time penod is a measurement of the amount of chitotπosidase protem present in a given volume of patient sample when compared agamst a standard curve A competition assay may be employed wherem antibodies specific to the chitotπosidase protem are attached to a sobd support and labeled protem and a sample deπved from the host are passed over the solid support and the amount of label detected attached to the solid support can be correlated to a quantity of chitotπosidase protem m the sample The following examples are provided for illustrative purposes only and are not intended to limit the invention. EXAMPLES Example 1 : Assay for smooth muscle migration Smooth muscle migration is measured in a chamber divided by a semi-permeable filter to which chitotriosidase (about 1 to 10 mg/ml) is bound on one side. On the other side, human fetal smooth muscle cells are cultured. The extent to which cells migrate into the filter is monitored colorimetrically (optical density). Chitotriosidase-induced migration response is measured aginst PDGF, a positive control and a known stimulator of smooth muscle migration. Example 2: Production and characterization of antibodies generated against chitotriosidase Murine monoclonal antibodies are produced using purified chitotriosidase. Purified chitotriosidase is used as the immunogen for a panel of mice (Charles Rive, Wilmington. MA). The animals receive three subcutaneous injections of chitotriosidase in phosphate buffered saline 0?BS) emulsified with a one to one ratio of TiterMAX® (CytoRx Corp., Norcross) over a period of four months. The priming antigen dose is about 50 mg and boosts are 25 mg and 10 mg. After boosts, serum samples are collected and assayed for binding to chitotriosidase. Animals producing serum samples that bind chitotriosidase are selected as spleen donors and boosted intravenously with 10 mg chitotriosidase prior to euthanasia. The fusion procedure, first reported by Kohler et al. (Nature 1975, 256, 495) is used with modifications to perform the technique using a cell monolayer (Kennet et al. Eds., "Hyb domas: A new dimension in biological analysis", pp. 368-377, Plenum Press, New York). Spleen cells from several, preferably two, donor mice are pooled and fusions performed using a ratio of 50 million spleen cells to ten million SP2/0/Agl4 myeloma cells. Supernatants from fusion-positive wells are assayed by binding to chitotriosidase by ELISA. Monoclonal antibodies are characterized for their ability to immunoprecipitate chitotriosidase with protein A sepharose. In addition, these monoclonal antibodies are tested for their ability to neutralize binding of chitotriosidase to sites found in tissue where chitotriosidase is expressed. Example 3: Expression of chitotriosidase in vitro A full-length cDNA has been subcloned into both mammalian and Drosophila expression vectors, CDN and MtaL ϋ. Expression of full-length protem is done in Drosophila S2 and CHO ACC317 cells. CHO Cell Transfection: A full length chitotπosidase gene fragment was isolated on a 1379 bp Eco Rl-Stu I fragment and ligated mto the CDN expression vector between Eco RI and Eco RV, thus plac g the transcπption unit under the control of the SMV promoter. Twenty micrograms of HC gp-39CDN plasmid construct was linearized by restnction digestion with Ssp I and electroporated into CHO ACC 317 (1.25 x IO 7 cells m 1 ml). Cells were seeded at a density of 2.5 x 10 3 cells per well and selected m minimal media in the absence of nucleosides. Drosophila S2 Transfection: A full-length chitotπosidase gene fragment was isolated on a 1403 bp Xba I-Stu I fragment and ligated mto the mtal JJ expression vector between Xba I and Stu I, thus placmg the transcπption unit under the control of the inducible Drosophila metallothionem promoter. 1.2 x IO 7 cells were cotransfected with 20 mg mtal ϋ-chitotπosidase chimenc construct and 1 mg pCOHygro plasmid conferring resistance to hygromycm. Stable lmes were selected in medium contammg 300 mg/ml hygromycm. SEQUENCE LISTING (1) GENERAL INFORMATION (i) APPLICANTS:SMITHKLINE BEECHAM CORPORATON AND HUMAN GENOME SCIENCES, INC. (ii) TITLE OF THE INVENTION: Chitotriosidase (iii) NUMBER OF SEQUENCES: 4 (iv) ' CORRESPONDENCE ADDRESS: (A) ADDRESSEE: SmithKline Beecham Corporation (B) STREET: 709 Swedeland Road (C) CITY: King of Prussia (D) STATE: PA (E) COUNTRY: USA (F) ZIP: 19406 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Diskette (B) COMPUTER: IBM Compatible (C) OPERATING SYSTEM: DOS (D) SOFTWARE: FastSEQ for Windows Version 2.0 ( i) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: Unknown (B) FILING DATE: Herein (C) CLASSIFICATION: Unknown ( ii) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: 60/014,295 (B) FILING DATE: 29-MAR-1996 (Viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Han, William T (B) REGISTRATION NUMBER: 34,344 (C) REFERENCE/DOCKET NUMBER: ATG50005 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: 610-270-5219 (B) TELEFAX: 610-270-4026 (C) TELEX: (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1637 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA (xi) SEQUENCE DESCRIPTION: SEQ ID Nθ:l: GGCACGAGCT ACCGCTGAGC TGCATCATGG TGCGGTCTGT GGCCTGGGCA GGTTTCATGG 60 TCCTGCTGAT GATCCCATGG GGCTCTGCTG CAAAACTGGT CTGCTACTTC ACCAACTGGG 120 CCCAGTACAG ACAGGGGGAG GCTCGCTTCC TGCCCAAGGA CTTGGACCCC AGCCTTTGCA 180 CCCACCTCAT CTACGCCTTC GCTGGCATGA CCAACCACCA GCTGAGCACC ACTGAGTGGA 240 ATGACGAGAC TCTCTACCAG GAGTTCAATG GCCTGAAGAA GATGAATCCC AAGCTGAAGA 300 CCCTGTTAGC CATCGGAGGC TGGAATTTCA GCACTCAGAA GTTCACAGAT ATGGTAGCCA 360 CGGCCAACAA CCGTCAGACC TTTGTCAACT CGGCCATCAG GTTTCTGCGC AAATACAGCT 420 TTGACGGCCT TGACCTTGAC TGGGAGTACC CAGGAAGCCA GGGGAGCCCT GCCGTAGACA 480 AGGAGCGCTT CACAACCCTG GACTTGGCCA ATGCCTTCCA GCAGGAAGCC CAGACCTCAG 540 GGAAGGAACG CCTTCTTCTG AGTGCAGCGG TTCCAGCTGG GCAGACCTAT GTGGATGCTG 600 GATACGAGGT GGACAAAATC GCCCAGAACC TGGATTTTGT CAACCTTATG GCCTACGACT 660 TCCATGGCTC TTGGGAGAAG GTCACGGGAC ATAACAGCCC CCTCTACAAG AGGCAAGAAG 720 AGAGTGGTGC AGCAGCCAGC CTCAACGTGG ATGCTGCTGT GCAACAGTGG CTGCAGAAGG 780 GGACCCCTGC CAGCAAGCTG ATCCTTGGCA TGCCTACCTA CGGACGCTCC TTCACACTGG 840 CCTCCTCATC AGACACCAGA GTGGGGGCCC CAGCCACAGG GTCTGGCACT CCAGGCCCCT 900 TCACCAAGGA AGGAGGGATG CTGGCCTACT ATGAAGTCTG CTCCTGGAAG GGGGCCACCA 960 AACAGAGAAT CCAGGATCAG AAGGTGCCCT ACATCTTCCG GGACAACCAG TGGGTGGGCT 1020 TTGATGATGT GGAGAGCTTC AAAACCAAGG TCAGCTATCT GAAGCAGAAG GGACTGGGCG 1080 GGGCCATGGT CTGGGCACTG GACTTAGATG ACTTTGCCGG CTTCTCCTGC AACCAGGGCC 1140 GATACCCCCT CATCCAGACG CTACGGCAGG AACTGAGTCT TCCATACTTG CCTTCAGGCA 1200 CCCCAGAGCT TGAAGTTCCA AAACCAGGTC AGCCCTCTGA ACCTGAGCAT GGTCCCAGCC 1260 CTGGACAAGA CACGTTCTGC CAGGGCAAAG CTGATGGGCT CTATCCCAAT CCTCGGGAAC 1320 GGTCCAGCTT CTACAGCTGT GCAGCGGGGC GGCTGTTCCA GCAAAGCTGC CCGACAGGCC 1380 TGGTGTTCAG CAACTCCTGC AAATGCTGCA CCTGGAATTG AGTCGCTAAA GCCCCTCCAG 1440 TCCCAGCTTT GAGGCTGGGC CCAGGATCAC TCTACAGCCT GCCTCCTGGG TTTTCCCTGG 1500 GGGCCGCAAT CTGGCTCCTG CAGGCCTTTC TGTGGTCTTC CTTTATCCAG GCTTTCTGCT 1560 CTCAGCCTTG CCTTCCTTTT TTCTGGGTCT CCTGGGCTGC CCCTTTCACT TGCAAAATAA 1620 ATCTTTGGTT TCTTCCC 1637 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 464 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Val Arg Ser Val Ala Trp Ala Gly Phe Met Val Leu Leu Met lie 1 5 10 15 Pro Trp Gly Ser Ala Ala Lys Leu Val Cys Tyr Phe Thr Asn Trp Ala 20 25 30 Gin Tyr Arg Gin Gly Glu Ala Arg Phe Leu Pro Lys Asp Leu Asp Pro 35 40 45 Ser Leu Cys Thr His Leu lie Tyr Ala Phe Ala Gly Met Thr Asn His 50 55 60 Gin Leu Ser Thr Thr Glu Trp Asn Asp Glu Thr Leu Tyr Gin Glu Phe 65 70 75 80 Asn Gly Leu Lys Lys Met Asn Pro Lys Leu Lys Thr Leu Leu Ala lie 85 90 95 Gly Gly Trp Asn Phe Ser Thr Gin Lys Phe Thr Asp Met Val Ala Thr 100 105 110 Ala Asn Asn Arg Gin Thr Phe Val Asn Ser Ala lie Arg Phe Leu Arg 115 120 125 Lys Tyr Ser Phe Asp Gly Leu Asp Leu Asp Trp Glu Tyr Pro Gly Ser 130 135 140 Gin Gly Ser Pro Ala Val Asp Lys Glu Arg Phe Thr Thr Leu Asp Leu 145 150 155 160 Ala Asn Ala Phe Gin Gin Glu Ala Gin Thr Ser Gly Lys Glu Arg Leu 165 170 175 Leu Leu Ser Ala Ala Val Pro Ala Gly Gin Thr Tyr Val Asp Ala Gly 180 185 190 Tyr Glu Val Asp Lys lie Ala Gin Asn Leu Asp Phe Val Asn Leu Met 195 200 205 Ala Tyr Asp Phe His Gly Ser Trp Glu Lys Val Thr Gly His Asn Ser 210 215 220 Pro Leu Tyr Lys Arg Gin Glu Glu Ser Gly Ala Ala Ala Ser Leu Asn 225 230 235 240 Val Asp Ala Ala Val Gin Gin Trp Leu Gin Lys Gly Thr Pro Ala Ser 245 250 255 Lys Leu lie Leu Gly Met Pro Thr Tyr Gly Arg Ser Phe Thr Leu Ala 260 265 270 Ser Ser Ser Asp Thr Arg Val Gly Ala Pro Ala Thr Gly Ser Gly Thr 275 280 285 Pro Gly Pro Phe Thr Lys Glu Gly Gly Met Leu Ala Tyr Tyr Glu Val 290 295 300 Cys Ser Trp Lys Gly Ala Thr Lys Gin Arg He Gin Asp Gin Lys Val 305 310 315 320 Pro Tyr He Phe Arg Asp Asn Gin Trp Val Gly Phe Asp Asp Val Glu 325 330 335 Ser Phe Lys Thr Lys Val Ser Tyr Leu Lys Gin Lys Gly Leu Gly Gly 340 345 350 Ala Met Val Trp Ala Leu Asp Leu Asp Asp Phe Ala Gly Phe Ser Cys 355 360 365 Asn Gin Gly Arg Tyr Pro Leu He Gin Thr Leu Arg Gin Glu Leu Ser 370 375 380 Leu Pro Tyr Leu Pro Ser Gly Thr Pro Glu Leu Glu Val Pro Lys Pro 385 390 395 400 Gly Gin Pro Ser Glu Pro Glu His Gly Pro Ser Pro Gly Gin Asp Thr 405 410 415 Phe Cys Gin Gly Lys Ala Asp Gly Leu Tyr Pro Asn Pro Arg Glu Arg 420 425 430 Ser Ser Phe Tyr Ser Cys Ala Ala Gly Arg Leu Phe Gin Gin Ser Cys 435 440 445 Pro Thr Gly Leu Val Phe Ser Asn Ser Cys Lys Cys Cys Thr Trp Asn 450 455 460 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1768 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: Other (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3 : CAAGCTCGAC ACTAACCCTC ACTAAAGGGA ACAAAAGCTG GAGCTCCACC GCGGTGCGGC 60 CGCTCTAGAA CTAGTGGATC CCCCGGGCTG CAGGAATTCG GCACGAGGCC GCTGAGCTGC 120 ATCATGGTGC GGTCTGTGGC CTGGGCAGGT TTCATGGTCC TGCTGATGAT CCCATGGGGC 180 TCTGCTGCAA AACTGGTCTG CTACTTCACC AACTGGGCCC AGTACAGACA GGGGGAGGCT 240 CGCTTCCTGC CCAAGGACTT GGACCCCAGC CTTTGCACCC ACCTCATCTA CGCCTTCGCT 300 GGCATGACCA ACCACCAGCT GAGCACCACT GAGTGGAATG ACGAGACTCT CTACCAGGAG 360 TTCAATGGCC TGAAGAAGAT GAATCCCAAG CTGAAGACCC TGTTAGCCAT CGGAGGCTGG 420 AATTTCRGCA CTCAGAAGTT CACAGATATG GTAGCCACGG CCAACAACCG TCAGACCTTT 480 GTCAACTCGG CCATCAGGTT TCTGCGCAAA TACAGCTTTG ACGGCCTTGA CCTTGACTGG 540 GAGTACCCAG GAAGCCAGGG GAGCCCTGCC GTAGACAAGG AGCGCTTCAC AACCCTGGTA 600 CAGGACTTGG CCAATGCCTT CCAGCAGGAA GCCCAGACCT CAGGGAAGGA ACGCCTTCTT 660 CTGAGTGCAG CGGTTCCAGC TGGGCAGACC TATGTGGATG CTGGATACGA GGTGGACAAA 720 ATCGCCCAGA ACCTGGATTT TGTCAACCTT ATGGCCTACG ACTTCCATGG CTCTTGGGAG 780 AAGGTCACGG GACATAACAG CCCCCTCTAC AAGAGGCAAG AAGAGAGTGG TGCAGCAGCC 840 AGCCTCAACG TGGATGCTGC TGTGCAACAG TGGCTGCAGA AGGGGACCCC TGCCAGCAAG 900 CTGATCCTTG GCATGCCTAC CTACGGACGC TCCTTCACAC TGGCCTCCTC ATCAGACACC 960 AGAGTGGGGG CCCCAGCCAC AGGGTCTGGC ACTCCAGGCC CCTTCACCAA GGAAGGAGGG 1020 ATGCTGGCCT ACTATGAAGT CTGCTCCTGG AAGGGGGCCA CCAAACAGAG AATCCAGGAT 1080 CAGAAGGTGC CCTACATCTT CCGGGACAAC CAGTGGGTGG GCTTTGATGA TGTGGAGAGC 1140 TTCAAAACCA AGGTCAGCTA TCTGAAGCAG AAGGGACTGG GCGGGGCCAT GGTCTGGGCA 1200 CTGGACTTAG ATGACTTTGC CGGCTTCTCC TGCAACCAGG GCCGATACCC CCTCATCCAG 1260 ACGCTACGGC AGGAACTGAG TCTTCCATAC TTGCCTTCAG GCACCCCAGA GCTTGAAGTT 1320 CCAAAACCAG GTCAGCCCTC TGAACCTGAG CATGGCCCCA GCCCTGGACA AGACACGTTC 1380 TGCCAGGGCA AAGCTGATGG GCTCTATCCC AATCCTCGGG AACGGTCCAG CTTCTACAGC 1440 TGTGCAGCGG GGCGGCTGTT CCAGCAAAGC TGCCCGACAG GCCTGGTGTT CAGCAACTCC 1500 TGCAAATGCT GCACCTGGAA TTGAGTCGCT AAAGCCCCTC CAGTCCCAGC TTTGAGGCTG 1560 GGCCCAGGAT CACTCTACAG CCTGCCTCCT GGGTTTTCCC TGGGGGCCGC AATCTGGCTC 1620 CTGCAGGCCT TTCTGTGGTC TTCCTTTATC CAGGCTTTCT GCTCTCAGCC TTGCCTTCCT 1680 TTTTTCTGGG TCTCCTGGGC TGCCCCTTTT ACTTGCAAAA TAAATCTTTG GTTTGTGCCC 1740 CTCTTCCCAA AAAAAAAAAA AAAAAAAA 1768 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1135 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Met Val Arg Ser Val Ala Trp Ala Gly Phe Met Val Leu Leu Met He 1 5 10 15 Pro Trp Gly Ser Ala Ala Lys Leu Val Cys Tyr Phe Thr Asn Trp Ala 20 25 30 Gin Tyr Arg Gin Gly Glu Ala Arg Phe Leu Pro Lys Asp Leu Asp Pro 35 40 45 Ser Leu Cys Thr His Leu He Tyr Ala Phe Ala Gly Met Thr Asn His 50 55 60 Gin Leu Ser Thr Thr Glu Trp Asn Asp Glu Thr Leu Tyr Gin Glu Phe 65 70 75 80 Asn Gly Leu Lys Lys Met Asn Pro Lys Leu Lys Thr Leu Leu Ala He 85 90 95 Gly Gly Trp Asn Phe Xaa Thr Gin Lys Phe Thr Asp Met Val Ala Thr 100 105 110 Ala Asn Asn Arg Gin Thr Phe Val Asn Ser Ala He Arg Phe Leu Arg 115 120 125 Lys Tyr Ser Phe Asp Gly Leu Asp Leu Asp Trp Glu Tyr Pro Gly Ser 130 135 140 Gin Gly Ser Pro Ala Val Asp Lys Glu Arg Phe Thr Thr Leu Val Gin 145 150 155 160 Asp Leu Ala Asn Ala Phe Gin Gin Glu Ala Gin Thr Ser Gly Lys Glu 165 170 175 Arg Leu Leu Leu Ser Ala Ala Val Pro Ala Gly Gin Thr Tyr Val Asp 180 185 190 Ala Gly Tyr Glu Val Asp Lys He Ala Gin Asn Leu Asp Phe Val Asn 195 200 205 Leu Met Ala Tyr Asp Phe His Gly Ser Trp Glu Lys Val Thr Gly His 210 215 220 Asn Ser Pro Leu Tyr Lys Arg Gin Glu Glu Ser Gly Ala Ala Ala Ser 225 230 235 240 Leu Asn Val Asp Ala Ala Val Gin Gin Trp Leu Gin Lys Gly Thr Pro 245 250 255 Ala Ser Lys Leu He Leu Gly Met Pro Thr Tyr Gly Arg Ser Phe Thr 260 265 270 Leu Ala Ser Ser Ser Asp Thr Arg Val Gly Ala Pro Ala Thr Gly Ser 275 280 285 Gly Thr Pro Gly Pro Phe Thr Lys Glu Gly Gly Met Leu Ala Tyr Tyr 290 295 300 Glu Val Cys Ser Trp Lys Gly Ala Thr Lys Gin Arg He Gin Asp Gin 305 310 315 320 Lys Val Pro Tyr He Phe Arg Asp Asn Gin Trp Val Gly Phe Asp Asp 325 330 335 Val Glu Ser Phe Lys Thr Lys Val Ser Tyr Leu Lys Gin Lys Gly Leu 340 345 350 Gly Gly Ala Met Val Trp Ala Leu Asp Leu Asp Asp Phe Ala Gly Phe 355 360 365 Ser Cys Asn Gin Gly Arg Tyr Pro Leu He Gin Thr Leu Arg Gin Glu 370 375 380 Leu Ser Leu Pro Tyr Leu Pro Ser Gly Thr Pro Glu Leu Glu Val Pro 385 390 395 400 Lys Pro Gly Gin Pro Ser Glu Pro Glu His Gly Pro Ser Pro Gly Gin 405 410 415 Asp Thr Phe Cys Gin Gly Lys Ala Asp Gly Leu Tyr Pro Asn Pro Arg 420 425 430 Glu Arg Ser Ser Phe Tyr Ser Cys Ala Ala Gly Arg Leu Phe Gin Gin 435 440 445 Ser Cys Pro Thr Gly Leu Val Phe Ser Asn Ser Cys Lys Cys Cys Thr 450 455 460 Trp Asn 465

Claims: What is claimed is 1 A nucleic acid sequence encodmg chitotnosidase 2 A method of diagnosing a tissue remodelmg disorder related to expression of a mutated chitotπosidase protein in a host compnsing carrying out nucleic acid amplification 3 A method of detecting altered expression of a chitotπosidase protein in a host compnsing contacting a bodily sample with antibody

Inventors: CHOPRA ARVIND, GENTZ REINER, KIRKPATRICK ROBERT B, NI JIAN, THOTAKURA NAGESWARA R, CHOPRA, ARVIND, GENTZ, REINER, KIRKPATRICK, ROBERT, B., NI, JIAN, THOTAKURA, NAGESWARA, R.
Applicants: CHOPRA ARVIND, GENTZ REINER, HUMAN GENOME SCIENCES INC, KIRKPATRICK ROBERT B, NI JIAN, SMITHKLINE BEECHAM CORP, THOTAKURA NAGESWARA R, CHOPRA, ARVIND, GENTZ, REINER, HUMAN GENOME SCIENCES, INC., KIRKPATRICK, ROBERT, B., NI, JIAN, SMITHKLINE BEECHAM CORPORATION, THOTAKURA, NAGESWARA, R.
Application year: 1997
Main classification: C07H21
IPC codes: C12N9, C12N15, G01N33