Platelet activating factor acetylhydrolase, and gene thereof

The present invention relates to an isolated human protein having platelet activating factor acetylhydrolase activity.

BACKGROUND OF THE INVENTION
 a) Field of the Invention
 This invention relates to a novel platelet activating factor
 acetylhydrolase, and a gene encoding the same.
 b) Description of the Related Art
 A platelet activating factor acetylhydrolase is an enzyme, which acts on a
 platelet activating factor (hereinafter abbreviated as "PAF") and
 eliminates its 2-acetyl group to deprive PAF of its activity. Since PAF is
 a mediator for inflammation which causes defluxion of tissue fluid through
 finer vessels, vasodilation, smooth muscle contraction, endothelial
 adhesion, activation of neutrophils, macrophages or eosinophilic
 leukocytes, or the like, PAF acetylhydrolase is usable as a preventive or
 therapeutic for various diseases caused by PAF.
 Some reports have been made about PAF acetylhydrolase to date. For its use
 as a medicine, however, there is an outstanding desire for the provision
 of a PAF acetylhydrolase having higher purity and stronger action compared
 with conventional PAF acetylhydrolase. Further, from the viewpoint of
 safety, PAF acetylhydrolase derived form human being instead of an animal
 is desired.
 SUMMARY OF THE INVENTION
 With the foregoing in view, the present invention has as a primary object
 the provision of PAF acetylhydrolase which can fulfill the above-described
 desires.
 Interested in the wide-spread distribution of PAF acetylhydrolase in animal
 organs such as the brain and kidneys, the present inventors chose the
 bovine liver as a source, and by various isolation and purification
 procedures, progressively increased the purity of PAF acetylhydrolase
 while placing a focus on its enzymatic activity. As a result, the present
 inventors have succeeded in obtaining bovine PAF acetylhydrolase as a pure
 product and further in determining its amino acid sequence. In addition,
 from the amino acid sequence of the PAF acetylhydrolase, a gene encoding
 the enzyme has been found by methods known per se in the art.
 Moreover, using the bovine PAF acetylhydrolase cDNA, the present inventors
 have also succeeded in identifying the human PAF acetylhydrolase cDNA.
 The present invention has been completed based on these findings, and
 provides a human PAF acetylhydrolase, which plays an important role as a
 PAF-inhibiting substance, and also a gene which encodes the enzyme and is
 important for the synthesis of the enzyme by genetic engineering.
 The human PAF acetylhydrolase according to the present invention
 selectively degrades PAF and hence, is usable as medicines or biochemical
 reagents for the prevention and treatment of diseases caused by PAF, for
 example, diseases such as asthma, exudative tympanitis, hemorrhagic
 colitis and adult respiratory distress syndrome.
 DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS
 The human PAF acetylhydrolase according to the present invention can be
 prepared as will be described next. PAF acetylhydrolase is first collected
 from an animal. From the PAF acetylhydrolase, the animal PAF
 acetylhydrolase cDNA is determined. Using the animal PAF acetylhydrolase
 cDNA, the human PAF acetylhydrolase cDNA is detected from a human gene
 library. The human PAF acetylhydrolase cDNA is inserted in an appropriate
 vector and then cultures in an adequate host organism, whereby the human
 PAF acetylhydrolase is obtained.
 Upon practice of the present invention, it is first necessary to obtain
 animal PAF acetylhydrolase from an organ of an animal such as the brain,
 liver or kidneys by purifying it through repetitions of known isolation
 and purification procedures while using PAF acetylhydrolase activity as an
 index. A description will hereinafter be made of a process for obtaining
 PAF acetylhydrolase by using a bovine liver as an example.
 As the bovine liver to be used as a source, one obtained from a bovine
 immediately after its slaughter is preferred.
 After the bovine liver is first washed with an appropriate buffer (for
 example, 10 mM Tris-HCl buffer containing 250 mM sucrose and 1 mM EDTA and
 having a pH of 7.4), it is homogenized with the same buffer. The
 homogenate is then centrifuged to obtain a soluble fraction.
 Making combined use of hydrophobic chromatography, ion exchange
 chromatography, adsorption chromatography, gel filtration chromatography
 and the like, the soluble fraction is purified until a single peak is
 observed by Mono Q FPLC, so that PAF acetyl hydrolase can be obtained.
 Incidentally, PAF acetylhydrolase activity which is used as an index for
 the selective collection of the PAF-acetylhydrolase-containing fraction
 can be determined, for example, by the method disclosed in Japanese Patent
 Application Laid-Open (Kokai) No. HEI 7-39373.
 With respect to the bovine PAF acetylhydrolase obtained in the
 above-described manner, its amino acid sequence was investigated by a
 method known per se in the art. As a result, the amino acid sequence has
 been found to be represented by the following formula (III) (SEQ ID NO:1):
 Met Gly Val Asn Gln Ser Val Ser Phe Pro Pro Val Thr Gly Pro His Leu Val Gly
 Cys Gly Asp Val Met Glu Gly Gln Ser Leu Gln Gly Ser Phe Phe Arg Leu Phe
 Tyr Pro Cys Gln Glu Ala Glu Glu Thr Ser Glu Gln Pro Leu Trp Ile Pro Arg
 Tyr Glu Tyr Cys Ala Gly Leu Ala Glu Tyr Leu Lys Phe Asn Lys Arg Trp Gly
 Gly Leu Leu Phe Asn Leu Gly Val Gly Ser Cys Arg Leu Pro Val Ser Trp Asn
 Gly Pro Phe Lys Thr Lys Asp Ser Gly Tyr Pro Leu Ile Ile Phe Ser His Gly
 Met Gly Ala Phe Arg Thr Val Tyr Ser Ala Phe Cys Met Glu Leu Ala Ser Arg
 Gly Phe Val Val Ala Val Pro Glu His Arg Asp Gly Ser Ala Ala Ala Thr Cys
 Phe Cys Lys Gln Thr Pro Glu Glu Asn Gln Pro Asp Asn Glu Ala Leu Lys Glu
 Glu Trp Ile Pro His Arg Gln Ile Glu Glu Gly Glu Lys Glu Phe Tyr Val Arg
 Asn Tyr Gln Val His Gln Arg Val Ser Glu Cys Val Arg Val Leu Lys Ile Leu
 Gln Glu Val Thr Ala Gly Gln Ala Val Leu Asn Ile Leu Pro Gly Gly Leu Asp
 Leu Met Thr Leu Lys Gly Gly Ile Asp Val Ser Arg Val Ala Val Met Gly His
 Ser Phe Gly Gly Ala Thr Ala Ile Leu Ala Leu Ala Lys Glu Met Gln Phe Arg
 Cys Ala Val Ala Leu Asp Ala Trp Met Phe Pro Leu Glu His Asp Phe Tyr Pro
 Thr Ala Arg Gly Pro Ile Phe Phe Ile Asn Ala Glu Lys Phe Gln Thr Val Glu
 Thr Val Asn Leu Met Lys Lys Ile Cys Asp Gln His His Gln Ser Arg Ile Ile
 Thr Val Leu Gly Ser Val His Arg Ser Leu Thr Asp Phe Val Phe Val Ala Gly
 Asn Trp Ile Ser Lys Phe Phe Ser Ser His Thr Arg Gly Ser Leu Asp Pro Tyr
 Glu Gly Gln Glu Thr Val Val Arg Ala Met Leu Ala Phe Leu Gln Lys His Leu
 Asp Leu Lys Glu Asp Tyr Asp Gln Trp Asn Ash Phe Ile Glu Gly Ile Gly Pro
 Ser Leu Thr Pro Gly Ala Pro His His Leu Ser Ser Leu (III)
 Further, from the peptide sequence of the bovine PAF acetylhydrolase of the
 formula (III), a gene encoding the enzyme was determined by a method known
 per se in the art. The gene (hereinafter called the "bovine PAF
 acetylhydrolase cDNA") has been found to be identified by the following
 formula (IV) (SEQ ID NO:2):
 GTCGACCCACGCGTCCGAGTTGACCGT
 CTGGGCTGTTTCTGAGGGTCAACGTGACTCGCCGTCAAGTTCAGCCACTGCCCAAGTCGT
 CGTTCAGTTCAGTTGGTTATGAG ATG GGG GTC AAC CAG TCT GTG AGC TTC CCA CCC GTC
 ACG GGA CCC CAC CTC GTA GGC TGT GGG GAT GTG ATG GAG GGT CAG AGC CTC CAG
 GGC AGC TTC TTT CGA CTG TTC TAC CCG TGC CAA GAG GCA GAG GAG ACC TCG GAG
 CAG CCC CTG TGG ATT CCC CGC TAT GAG TAC TGC GCT GGC CTG GCC GAA TAC CTA
 AAG TTT AAT AAG CGC TGG GGG GGG TTA CTG TTC AAC CTG GGT GTG GGA TCT TGT
 CGC CTG CCT GTT AGC TGG AAT GGC CCC TTT AAA ACA AAG GAC TCT GGA TAC CCC
 TTG ATC ATC TTC TCT CAT GGC ATG GGA GCC TTC AGG ACA GTG TAT TCA GCC TTC
 TGC ATG GAG CTG GCT TCT CGT GGC TTT GTG GTT GCT GTA CCA GAG CAC AGG GAT
 GGG TCA GCT GCG GCC ACC TGT TTC TGC AAG CAG ACC CCA GAG GAG AAC CAG CCT
 GAC AAT GAG GCC CTG AAG GAG GAA TGG ATC CCC CAC CGT CAA ATT GAG GAA GGG
 GAG AAG GAA TTC TAT GTT CGG AAC TAC CAG GTG CAT CAG AGG GTG AGC GAG TGT
 GTG AGG GTG TTG AAG ATC CTA CAA GAG GTC ACT GCT GGG CAG GCC GTT CTC AAC
 ATC TTG CCT GGC GGA TTG GAT CTG ATG ACC TTG AAG GGC GGC ATT GAC GTG AGC
 CGT GTG GCT GTA ATG GGA CAT TCA TTT GGA GGG GCC ACA GCT ATT CTG GCC TTG
 GCC AAG GAG ATG CAA TTT AGG TGT GCT GTG GCT TTG GAC GCT TGG ATG TTT CCT
 CTG GAG CAT GAC TTT TAC CCC ACG GCC CGA GGC CCT ATC TTC TTT ATC AAT GCT
 GAG AAG TTC CAG ACA GTG GAG ACT GTC AAC TTG ATG AAA AAG ATT TGT GAC CAG
 CAC CAC CAA TCC AGG ATC ATA ACT GTC CTT GGT TCT GTT CAT CGG AGT CTA ACC
 GAC TTT GTT TTT GTG GCT GGT AAC TGG ATT AGT AAA TTC TTC TCC AGT CAC ACC
 CGT GGA AGC TTG GAC CCC TAT GAA GGT CAG GAG ACC GTG GTG CGG GCC ATG TTG
 GCC TTC CTG CAG AAG CAT CTT GAC CTG AAA GAG GAC TAT GAC CAG TGG AAC AAC
 TTC ATT GAA GGC ATT GGC CCA TCA CTG ACC CCA GGG GCC CCA CAC CAT CTG TCC
 AGC CTG TAG GCACAACTGGTCATCTTGTGGAAG
 GTCCCTGAGCTGAGTTCCCGTGTGGGGCCTGCCCAGGGATACCCTTGGCCTCCTATCAGG
 AAGTGATTGCCATGACCCTTCTGTGTTGATTGAGAGGATATAATCACACTGCTGATTGGT
 AACGGGGTACTTGGATTCTCAGACTTGTCGATCTTAAACTCATGTTGGGACTTGGGTTCA
 CTTACTGATGGGCAAACGGGCATTCTGAGGACTGAGCCTTAATGGTATGGAGAACAAACA
 GTGGGATGGGGCTGGGGAAGATCTAAGCCCTAAGCTGGGCACTATGAGCCCTATAAACCC
 AACCAGCCAACACCCTCACCTTGGGCAAGTATGACTTCTGCAGGTCGACTCT (IV)
 To obtain human PAF acetylhydrolase from the bovine PAF acetylhydrolase
 CDNA obtained as described above, the human gene library is screened by a
 method known per se in the art while using the bovine PAF acetylhydrolase
 CDNA as a template.
 Described specifically, the bovine PAF acetylhydrolase cDNA is labeled, for
 example, by incorporating fluorescein-12-dUTP through PCR. By the colony
 hybridization technique that selects each positive colony by ECL (Enhanced
 Chemiluminescence; Amersham K. K.), colonies containing the human PAF
 acetylhydrolase cDNA can be obtained.
 The human PAF acetylhydrolase cDNA obtained as described above has been
 found to be identified by the following formula (II) (SEQ ID NO:4):
 GCAGGTCTCGACCCACGCGTCCGCGGACGCGTGGG
 CGAGAAGTGCTTCCAAGCGTCCATTTTGAGCCTTGGAAACTACGACGACCAAAGGGCCAC
 GGGTTCCTGGGTCGTTTCTCATTTCCGTCGAGTTAAACGTCTGGGGCTGCTTCTGAGGAA
 TCAGCTTGGCTGGCCAGCAAGTTCAGCTCCGGCAAGTCATTTGATTCACCCGGTGATGAA ATG GGG GTC
 AAC CAG TCT GTG GGC TTT CCA CCT GTC ACA GGA CCC CAC CTC GTA GGC TGT GGG
 GAT GTG ATG GAG GGT CAG AAT CTC CAG GGG AGC TTC TTT CGA CTC TTC TAC CCC
 TGC CAA AAG GCA GAG GAG ACC ATG GAG CAG CCC CTG TGG ATT CCC CGC TAT GAG
 TAC TGC ACT GGC CTG GCC GAG TAC CTG CAG TTT AAT AAG CGC TGC GGG GGC TTG
 CTG TTC AAC CTG GCG GTG GGA TCT TGT CGC CTG CCT GTT AGC TGG AAT GGC CCC
 TTT AAG ACA AAG GAC TCT GGA TAC CCC TTG ATC ATC TTC TCC CAT GGC CTA GGA
 GCC TTC AGG ACT TTG TAT TCA GCC TTC TGC ATG GAG CTG GCC TCA CGT GGC TTT
 GTG GTT GCT GTG CCA GAG CAC AGG GAC CGG TCA GCG GCA ACC ACC TAT TTC TGC
 AAG CAG GCC CCA GAA GAG AAC CAG CCC ACC AAT GAA TCG CTG CAG GAG GAA TGG
 ATC CCT TTC CGT CGA GTT GAG GAA GGG GAG AAG GAA TTT CAT GTT CGG AAT CCC
 CAG GTG CAT CAG CGG GTA AGC GAG TGT TTA CGG GTG TTG AAG ATC CTG CAA GAG
 GTC ACT GCT GGG CAG ACT GTC TTC AAC ATC TTG CCT GGT GGC TTG GAT CTG ATG
 ACT TTG AAG GGC AAC ATT GAC ATG AGC CGT GTG GCT GTG ATG GGA CAT TCA TTT
 GGA GGG GCC ACA GCT ATT CTG GCT TTG GCC AAG GAG ACC CAA TTT CGG TGT GCG
 GTG GCT CTG GAT GCT TGG ATG TTT CCT CTG GAA CGT GAC TTT TAC CCC AAG GCC
 CGA GGA CCT GTG TTC TTT ATC AAT ACT GAG AAA TTC CAG ACA ATG GAG AGT GTC
 AAT TTG ATG AAG AAG ATA TGT GCC CAG CAT GAA CAG TCT AGG ATC ATA ACC GTT
 CTT GGT TCT GTT CAT CGG AGT CAA ACT GAC TTT GCT TTT GTG ACT GGC AAC TTG
 ATT GGT AAA TTC TTC TCC ACT GAA ACC CGT GGG AGC CTG GAC CCC TAT GAA GGG
 CAG GAG GTT ATG GTA CGG GCC ATG TTG GCC TTC CTG CAG AAG CAC CTC GAC CTG
 AAA GAA GAC TAT AAT CAA TGG AAC AAC CTT ATT GAA GGC ATT GGA CCG TCG CTC
 ACC CCA GGG GCC CCC CAC CAT CTG TCC AGC CTG TAG
 GCACAACTGGCCATTTGTAAAGTCACTTCAGCCAAGTTTTCATTTGGG
 AGCTACCCAAGGGCACCCATGAGCTCCTATCAAGAAGTGATCAACGTGACCCCTTTTCAC
 AGATTGAAAGGTGTAATCACACTGCTGCTTGGATAACTGGGTACTTTGATCTTAGATTTG
 ATCTTAAAATCACTTTGGGACTGGGATCCCTTGCTGATTGACAAACAGACTTTCTGGGAC
 CTTGATGGAGTGGGGAACAAGCAGTAGAGTGGGACTGGGGGAGACCCAGGCCCCGGGCTG
 AGCACTGTGAGGCCTGGATGTGAAGACTCAGCCCAGCGAAGCTCATTCCCTTACCCCCGG
 CCAGTGCTGCTGCTTCAGTGGAAGAGATGAAGCCAAAGGACAGAATGAAAATCCCTACCT
 TCAGAGACTCTAGCCCAGCCCAACACCATCTCTTCCTACCTCTCAGCCTTCTCCCTCCCC
 AGGGCCACTTGTTGAAGTCTGAGCACTTTATGTAAATTTCTAGGTGTGAGCCGTGATCAC
 ATTTTCTATTTATTTCCAAGTCTTCTCATTGTATGGAACATAGTACTACTTATACTTACA
 GTAGTAAGTTATACTTGTGAGCCCACAGAGTGGCAGACAGCATGGCTCTCACAGCACAGG
 GAGAAAAACTGAGGTACACAGAGGTACCTCAGAAGCTCTGGATGTCTTTGGGGGTTTTGC
 TAAGTGTATCTTGATAGGAAACAACAAAAGCAGGTTGAGATGGGGAAGATGACAGAACAA
 CAGTGTTAAATGGCCATTTGCACAGGCCTTTGCCACAACAGAGAAGTAGTTTGGTCAGCT
 AAAACTCAGCTGCAGCCTGGACAGTAGAGCGAGACCCCATCTTAAAAATAAAGAAGGCTG
 GGCGTGGTGGCTCATGCCTGTAATCCCAGCACTTTGGGAGGCCAAGGCAGGCAGATCACT
 TAAGGCCAGGAGTTCAAGACCACCTGGCCAACATGGTGAAACCCCGTCTCTACTAAAAAT
 ACAAAAAATTAGCCTGGCGTAATGGCAGGCGCCTATAATCCCAGCTACTCAGGAGGCTGA
 AGCAGAAGAATCACTTGAACCTAGGAGGCGGAGGTTGCAGTGAGTCAAGATCGCGCCACT
 GCACTCCAGCCTGGGTGACAGAGCAAGACTCTGTCTT (II)
 Following conventional procedures, the human PAF acetylhydrolase cDNA
 obtained as described above is next introduced in an appropriate vector
 plasmid, and host cells such as mammal cells are then transformed by a
 commonly-employed recombinant DNA technique to express the human PAF
 acetylhydrolase. The expression of the human PAF acetylhydrolase can be
 confirmed by a western blot technique which makes use of an anti-human PAF
 acetylhydrolase antibody. The introduction into the plasmid, the
 establishment of the transformed strain, the culture of the strain and the
 like can be conducted by the general recombinant DNA technology.
 From expression systems known to artisans, a suitable expression system can
 be selected for use in the present invention. It is possible to improve
 the efficiency of secretion and the level of expression by adding or
 improving a signal sequence and/or choosing an appropriate host. Although
 no particular limitation is imposed on host cells, illustrative examples
 include cultured cells of bacteria, yeasts, other fungi, human and other
 animals, and cultured cells of plants. Namely, the polynucleotide
 according to the present invention is inserted in a suitable expression
 vector, for example, pUC-PL-cl vector, the expression vector is introduced
 in adequate host cells, for example, E. Coli W3110 or the like, and the
 host cells are then cultured. The target human PAF acetylhydrolase can
 thereafter be coIlected as a protein from the thus-obtained cultured
 matter (cells or culture medium).
 As the host, a procaryote or an eucaryote can be used. Usable examples of
 the procaryote include bacteria, especially Escherichia coli and Bacillus
 bacteria, for example, B. subtilis. On the other hand, usable examples of
 the eucaryote include eucaryotic microorganisms such as yeasts, for
 example, Saccharomyces yeasts, especially S. Servisiae; insect cells such
 as armyworm (Spodoptera Frugiperda) cells and silkworm (Bombyx mori)
 cells; and animal cells such as human cells, monkey cells and mouse cells,
 especially monkey cells, for example, COS1 and COS 7.
 Usable examples of the expression vector include plasmids, pharges,
 phargemids, viruses [baculoviruses (for insect cells), vaccinia viruses
 (for animal cells)]. The promoter in the expression vector is selected
 depending on the host cells. For examples, lac promoters, trp promoters,
 trc promoters and the like can be used as promoters for bacteria; and adh
 1 promoters, pgk promoters and the like can be used as promoters for
 yeasts. Further, baculovirus polyhedrin promoters can be mentioned as
 promoters for insects; and early and late promoters of Simian virus 40
 (SV40) can be mentioned as promoters for animal cells.
 When an enhancer is used, for example, the enhancer of SV40 is inserted
 either upstream or downstream of the gene.
 The transformation of the host by the expression vector can be conducted by
 a common method known per se in the art. Such methods are disclosed, for
 example, in "Current Protocols in Molecular Biology", John Wiley & Sons,
 Inc.
 The culture of the transformants can also be conducted by a usual method.
 The purification of the human PAF acetylhydrolase from the cultured matter
 can be conducted following procedures commonly employed for the isolation
 and purification of proteins, for example, by ultrafiltration and/or one
 or more of various column chromatographic procedures, for example,
 chromatography making use of "Sepharose".
 In the above-described manner, the human PAF acetylhydrolase can be
 advantageously obtained. The human PAF acetylhydrolase according to the
 present invention is represented by the following formula (I) (SEQ ID
 NO:3):
 Met Gly Val Asn Gln Ser Val Gly Phe Pro Pro Val Thr Gly Pro His Leu Val Gly
 Cys Gly Asp Val Met Glu Gly Gln Asn Leu Gln Gly Ser Phe Phe Arg Leu Phe
 Tyr Pro Cys Gln Lys Ala Glu Glu Thr Met Glu Gln Pro Leu Trp Ile Pro Arg
 Tyr Glu Tyr Cys Thr Gly Leu Ala Glu Tyr Leu Gln Phe Asn Lys Arg Cys Gly
 Gly Leu Leu Phe Asn Leu Ala Val Gly Ser Cys Arg Leu Pro Val Ser Trp Asn
 Gly Pro Phe Lys Thr Lys Asp Ser Gly Tyr Pro Leu Ile Ile Phe Ser His Gly
 Leu Gly Ala Phe Arg Thr Leu Tyr Ser Ala Phe Cys Met Glu Leu Ala Ser Arg
 Gly Phe Val Val Ala Val Pro Glu His Arg Asp Arg Ser Ala Ala Thr Thr Tyr
 Phe Cys Lys Gln Ala Pro Glu Glu Asn Gln Pro Thr Asn Glu Ser Leu Gln Glu
 Glu Trp Ile Pro Phe Arg Arg Val Glu Glu Gly Glu Lys Glu Phe His Val Arg
 Asn Pro Gln Val His Gln Arg Val Ser Glu Cys Leu Arg Val Leu Lys Ile Leu
 Gln Glu Val Thr Ala Gly Gln Thr Val Phe Asn Ile Leu Pro GIy Gly Leu Asp
 Leu Met Thr Leu Lys Gly Asn Ile Asp Met Ser Arg Val Ala Val Met Gly His
 Ser Phe Gly Gly Ala Thr Ala Ile Leu Ala Leu Ala Lys Glu Thr Gln Phe Arg
 Cys Ala Val Ala Leu Asp Ala Trp Met Phe Pro Leu Glu Arg Asp Phe Tyr Pro
 Lys Ala Arg Gly Pro Val Phe Phe Ile Asn Thr Glu Lys Phe Gln Thr Met Glu
 Ser Val Asn Leu Met Lys Lys Ile Cys Ala Gln His Glu Gln Ser Arg Ile Ile
 Thr Val Leu Gly Ser Val His Arg Ser Gln Thr Asp Phe Ala Phe Val Thr Gly
 Asn Leu Ile Gly Lys PIle Phe Ser Thr Glu Thr Arg Gly Ser Leu Asp Pro Tyr
 Glu Gly Gln Glu Val Met Val Arg Ala Met Leu Ala Phe Leu Gln Lys His Leu
 Asp Leu Lys Glu Asp Tyr Asn Gln Trp Asn Asn Leu Ile Glu Gly Ile GIy Pro
 Ser Leu Thr Pro Gly Ala Pro His His Leu Ser Ser Leu (I)
 The human PAF acetylhydrolase selectively degrades PAF and oxidized
 phospholipids and has physiologically active effects such
 anti-inflammatory effects.
 Needless to say, the human PAF acetylhydrolase according to the present
 invention is not limited to the peptide of the formula (I) (SEQ ID NO:3)
 but includes peptides having homology therewith, namely, peptides having
 the same function as the peptide represented by the formula (I) (SEQ ID
 NO:3) despite substitution, deletion, addition or the like of amino acids
 at parts of their sequences.
 The bovine PAF acetylhydrolase represented by the formula (III) (SEQ ID
 NO:4) may be contemplated to be available by gene manipulation in a
 similar manner as the human PAF acetylhydrolase. As a matter of fact,
 however, the bovine PAF acetylhydrolase cannot be obtained unless
 eucaryotic host cells are used.
 To obtain the bovine PAF acetylhydrolase by gene manipulation, it is
 therefore necessary to employ as host cells those derived from an
 eucaryote and to select and use a vector compatible with the host cells.
 An antibody against the human PAF acetylhydrolase or bovine PAF
 acetylhydrolase (which may hereinafter be collectively called the "PAF
 acetylhydrolase") according to the present invention can also be obtained
 following usual procedures.
 Described specifically, the antibody can be obtained by sensitizing an
 animal such as a rabbit with the PAF acetylhydrolase, separating its serum
 and, if necessary, purifying an immunoglobulin fraction from the serum. To
 enhance the sensitizing ability of the enzyme in this case, the enzyme in
 a form bound on a carrier protein such as bovine serum albumin (BSA) or
 methyl BSA may be used as an immunogen.
 Upon sensitizing an animal, the enzyme can also be used together with
 Freund's complete adjuvant (FCA) or Freund's incomplete adjuvant (FICA) to
 increase the production of the antibody. It is desired to conduct the
 sensitization of the animal twice or more. The frequency of sensitization
 can be determined while checking the antibody titer of the serum by test
 sampling of blood. The whole blood of an immune animal may be used by
 slaughtering it as needed. As an alternative, an immune animal may be
 subjected to booster sensitization as many times as needed to maintain a
 constant antibody titer, and blood samples may be collected in small
 quantities as needed for immediate use. It is also possible to obtain a
 monoclonal antibody in a usual manner by sensitizing a mouse with the
 enzyme and then forming hybridomas from spleen cells and myeloma cells of
 the sensitized mouse.

The present invention will hereinafter be described in further detail by
 the following examples and reference examples. It is however to be noted
 that the present invention are by no means limited by or to these
 examples.
 REFERENTIAL EXAMPLE 1
 Measurement of PAF Acetylhydrolase Activity
 (1) Using unlabeled lyso PAF (product of Bachem Feinchemikalien AG),
 1-O-[1-.sup.14 C]hexadecyl-lyso PAF (product of New England Nuclear
 Company; hereinafter called the "labeled lyso PAF") was diluted to 4,000
 dpm/nmol.
 On the other hand, 1-O-hexadecyl-2-[.sup.3
 H-acetyl]-sn-glycero-3-phosphocholine (hereinafter called ".sup.3 H-acetyl
 PAF") was diluted to 3,200 dpm/nmol with the unlabeled lyso PAF.
 A standard culture system for the measurement of PAF acetylhydrolase was
 composed of 50 mM Tris-HCl (pH 7.4), 5 mM EDTA, 5 mM 2-mercaptoethanol
 (2-ME) and 20 nmol .sup.3 H-acetyl PAF. The total volume of the sample was
 0.25 ml.
 (2) Measurement of PAF acetylhydrolase activity was conducted by culturing
 a test sample in the above-described standard culture system at 37.degree.
 C. for 30 minutes, adding 2.5 ml of chloroform/methanol (4:1 V/V) and 0.25
 ml of water to terminate the reaction, and then measuring the
 radioactivity of a small amount (0.6 ml) of each upper layer to determine
 the amount of the acetate liberated from the .sup.3 H-acetyl PAF.
 EXAMPLE 1
 Obtainment of Bovine PAF Acetylhydrolase
 (1) A fresh bovine liver was purchased from a slaughterhouse and was then
 treated within 3 hours of the slaughter. Treatments were all conducted at
 0 to 4.degree. C. The liver was homogenized in a Waring blender subsequent
 to the addition of a homogenizing buffer [10 mM Tris-HCl (pH 7.4), 250 mM
 sucrose, 1 mM EDTA] in an amount 5 times as much as the liver. The
 resulting homogenate was centrifuged for 30 minutes under 100,000.times.g,
 followed by the removal of a solid portion. The resultant supernatant was
 centrifuged further for 1 hour under 100,000.times.g, whereby a dissolved
 portion was obtained (supernatant portion)
 (2) The supernatant portion obtained through the procedures (1) was
 adjusted to 1 M with NaCl. Subsequent to stirring for 15 minutes, the
 solution was loaded on a "BUTYL TOYOPEARL 650 M" column which had been
 equilibrated beforehand with a buffer composed of 50 mM Tris-HCl (pH 7.4),
 1 mM EDTA and 1 M NaCl. After the column was washed with the same buffer,
 proteins were eluted with a linear gradient of NaCl (1 to 0 M). PAF
 acetylhydrolase activity was eluted as a single peak in 1 to 0 M NaCl
 fractions.
 (3) Active fractions from the "BUTYL TOYOPEARL" column were loaded on a
 "Q-Sepharosel" column which had been equilibrated with 10 mM Tris-HCl (pH
 7.4), 1 mM EDTA and 20% (V/V) glycerol (buffer A). The column was washed
 with the buffer A. Proteins were eluted with a linear gradient of NaCl (0
 to 500 mM) in the buffer A. The activity was observed in a fraction eluted
 with about 300 mM NaCl.
 (4) The active fraction from the "Q-Sepharose" column was concentrated to
 about 6 ml in an "Amicon ultrafiltration cell" in which "YM-10" membranes
 were used. The thus-concentrated fraction was loaded on a "Biogel A-1.5 m"
 gel filtration column which had been equilibrated beforehand with 10 mM
 Tris-HCl (pH 7.4), 200 mM NaCl, 5 mM 2-ME, 20% (V/V) glycerol and 0.5%
 (W/V) "CHAPS" (buffer B). The activity was eluted as a single peak in a
 fraction corresponding to a molecular weight of about 40 kDa.
 (5) The active fraction from the "Bioge1-A 1.5 m" column was loaded on a
 hydroxyapatite column which had been equilibrated beforehand with 10 mM
 Tris-HCl (pH 7.4), 5 mM 2-ME, 20% (V/V) glycerol and 0.5% (W/V) "CHAPS"
 (buffer C). Proteins were eluted with a linear gradient which ranged from
 the buffer C alone to a buffer C containing 150 mM KH.sub.2 PO.sub.4. The
 activity was observed in a fraction which was eluted with about 50 mM
 KH.sub.2 PO.sub.4.
 (6) The active fraction from the hydroxyapatite column was dialyzed against
 the buffer C, and was then loaded on an "FPLC Mono Q HR 5/5" column which
 had been equilibrated beforehand with the buffer C. Proteins were eluted
 by a linear gradient of NaCl (0 to 500 mM) in the buffer C. The activity
 was observed in a fraction which was eluted with 250 mM NaCl, and a
 protein in the fraction was obtained as purified bovine PAF
 acetylhydrolase.
 The total proteins, total activities, purification degrees (in terms of
 times) and the like in the individual purification steps described above
 are tabulated below:

Activity Degree of
 Total proteins Total activity per weight purification
 Yield
 Step (mg) (.mu.mol/min) (nmon/min/mg) (times)
 (%)
 Cytoplasm 46000 73.5 1.6 1 100
 BUTYL TOYOPEAL 680 16.3 24 15 22
 Q Sepharose FF 72.4 8.96 124 78 12
 Biogel A-1.5 m 6.93 7.38 1060 670 10
 Hydroxyapatite 3.45 5.29 1530 960
 7.2
 Mono Q FPLC 0.3 2.16 7200 4500
 2.9
 EXAMPLE 2
 Determination of Amino Acid Sequence of Bovine PAF Acetylhydrolase
 (1) About 0.2 mg of the purified PAF acetylhydrolase obtained in Example 1
 was reduced with 1 mg of dithiothreitol at room temperature for 2 hours,
 followed by the S-alkylation with 0.6% (W/V) 4-vinylpyridine at room
 temperature for 2 hours.
 Using a 4.6 mm.times.250 mm "Vydak 304-1251 C.sub.4 " column which had been
 equilibrated beforehand with 20% (V/V) acetonitrile containing 0.1% (V/V)
 trifluoroacetic acid, the reaction mixture was subjected to reverse phase
 high-performance liquid chromatography (HPLC). Proteins were then eluted
 with a linear gradient of acetonitrile (20 to 85% V/V) which contained
 0.1% (V/V) trifluoroacetic acid.
 (2) 40 kDa polypeptide, which had been purified by the HPLC, was dialyzed
 against a lysylendopeptidase digestive buffer [0.5 M Tris-HCl (pH 8.5) and
 4 M urea]. Next, 1 .mu.g of a lysylendopeptidase was added to the sample.
 After the reaction mixture was incubated for 18 hours at 37.degree. C.,
 the reaction mixture was fractionated by reverse phase HPLC through a 4.6
 mm.times.250 mm "Vydak 304-1251 C.sub.4 " column while using a linear
 gradient of acetonitrile (5 to 70% V/V) which contained 0.1% (V/V)
 trifluoroacetic acid.
 (3) The amino acid sequence of a peptide fragment obtained by the reverse
 phase HPLC was determined by an automated sequencer ("Model 477A", trade
 name; manufactured by AppIled Biosystems, Inc.).
 The base sequence of the bovine PAF acetylhydrolase, which was determined
 from the amino acid sequence of the peptide fragment, was as shown above
 by the formula (III) (SEQ ID NO:4).
 Further, from the peptide sequence (III) (SEQ ID NO:1)of the bovine PAF
 acetylhydrolase, a gene encoding the enzyme was determined by a method
 known per se in the art. The gene was found to be represented by the
 formula (IV) (SEQ ID NO:2).
 EXAMPLE 3
 Cloning of Non-active Human PAF Acetylhydrolase CDNA
 Using as a template the bovine PAF acetylhydrolase cDNA obtained in Example
 2, fluorescein-12-dUTP was incorporated in 500,000 clones of each of a
 fetal human liver cDNA library (pRc/CMV) and a human brain cDNA library
 (pCMV SPORTS) by PCR. The clones were then subjected to colony
 hybridization while detecting the labeling reagent by ECL, whereby cloning
 was conducted. As a result, a single positive clone was obtained from the
 human brain library.
 A plasmid DNA was prepared and the base sequence was determined. The clone
 was a full-length clone which contained ATG encoding initiating
 methionine. Encoding 43 N-terminal amino acids were the same as the
 corresponding amino acids in the sequence of the bovine PAF
 acetylhydrolase up to the 40th amino acid, and there was poly A at the 3'
 end. A more accurate determination of the base sequence was conducted. As
 a result, the cDNA was found to consist of 2188 bp and to contain an ORF
 (open reading frame) consisting of 253 amino acids. Compared with the
 bovine PAF acetylhydrolase cDNA, 140 amino acids had been deleted. The
 segment of the deleted 140 amino acids contains a "catalytic triad" of
 serine, histidine and aspartic acid, which exhibits catalytic activity.
 The cDNA is therefore not believed to have PAD acetylhydrolase activity.
 Hence, a primer was synthesized at positions flanking the deleted region,
 and PCR was conducted using the library DNA as a template. From the human
 brain cDNA, two bands were obtained, one corresponding to the
 above-described cDNA with the 140 amino acids deleted, and the other to a
 cDNA having substantially the same length as the bovine PAF
 acetylhydrolase cDNA. From the foregoing, the human brain library DNA was
 expected to contain, in addition to the above-obtained cDNA, a human PAF
 acetylhydrolase CDNA which is actually equipped with PAF acetylhydrolase
 activity.
 EXAMPLE 4
 Cloning of Human PAF Acetylhydrolase cDNA
 The human brain cDNA library was diluted to give 2000 clones per well,
 followed by incubation on 5 96-well plates. Subpools consisting of 10
 wells were prepared, and positive pools were determined by PCR (Pool Nos.
 10, 20, 28, and 38). With respect to these subpools, PCR was conducted
 well after well, so that positive pools were confirmed (Pool Nos. 10-5,
 20-10, and 38-12).
 Concerning these pools, incubation was conducted on plates subsequent to
 dilution. Using the non-active human PAF acetylhydrolase cDNA as a probe,
 cloning was attempted by hybridization. Labeling of the DNA was conducted
 with fluorescein 12-dUTP by PCR, and detection was carried out by ECL.
 Positive colonies were obtained from Pool Nos. 10-5 and 20-10. Plasmid
 DNAs of these clones were replicated, and their base sequences were then
 determined. As a result, a human PAF acetylhydrolase cDNA represented by
 the formula (II) (SEQ ID NO:4) was obtained from the clones of Pool Nos.
 10-5.
 Based on the resultant cDNA, the amino acid sequence of the human PAF
 acetylhydrolase was determined. It was found to be represented by the
 formula (I) (SEQ ID NO:3). Up to 88%, the sequence was the same as that of
 the bovine PAF acetylhydrolase (346/392 amino acids). On the other hand,
 it was 42% identical to that of the plasma human PAF acetylhydrolase
 (162/392 amino acids).
 Further, the above cDNA was incorporated in the pUC-Pl-cl vector,
 introduced in E. coli W3110 and then subjected to expression. A band,
 which corresponded to a protein having a molecular weight of 42 kDa, was
 detected by SDS-PAGE.
 The protein was investigated for activity. Human PAF acetylhydrolase
 activity was confirmed.
 SEQUENCE LISTING
 (1) GENERAL INFORMATION:
 (iii) NUMBER OF SEQUENCES: 4
 (2) INFORMATION FOR SEQ ID NO: 1:
 (i) SEQUENCE CHARACTERISTICS:
 (A) LENGTH: 392 amino acids
 (B) TYPE: amino acid
 (C) STRANDEDNESS: single
 (D) TOPOLOGY: linear
 (ii) MOLECULE TYPE: peptide
 (vi) ORIGINAL SOURCE:
 (A) ORGANISM: BOVINE (Bos taurus)
 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1
 Met Gly Val Asn Gln Ser Val Ser Phe Pro Pro Val Thr Gly Pro His
 1 5 10 15
 Leu Val Gly Cys Gly Asp Val Met Glu Gly Gln Ser Leu Gln Gly Ser
 20 25 30
 Phe Phe Arg Leu Phe Tyr Pro Cys Gln Glu Ala Glu Glu Thr Ser Glu
 35 40 45
 Gln Pro Leu Trp Ile Pro Arg Tyr Glu Tyr Cys Ala Gly Leu Ala Glu
 50 55 60
 Tyr Leu Lys Phe Asn Lys Arg Trp Gly Gly Leu Leu Phe Asn Leu Gly
 65 70 75 80
 Val Gly Ser Cys Arg Leu Pro Val Ser Trp Asn Gly Pro Phe Lys Thr
 85 90 95
 Lys Asp Ser Gly Tyr Pro Leu Ile Ile Phe Ser His Gly Met Gly Ala
 100 105 110
 Phe Arg Thr Val Tyr Ser Ala Phe Cys Met Glu Leu Ala Ser Arg Gly
 115 120 125
 Phe Val Val Ala Val Pro Glu His Arg Asp Gly Ser Ala Ala Ala Thr
 130 135 140
 Cys Phe Cys Lys Gln Thr Pro Glu Glu Asn Gln Pro Asp Asn Glu Ala
 145 150 155 160
 Leu Lys Glu Glu Trp Ile Pro His Arg Gln Ile Glu Glu Gly Glu Lys
 165 170 175
 Glu Phe Tyr Val Arg Asn Tyr Gln Val His Gln Arg Val Ser Glu Cys
 180 185 190
 Val Arg Val Leu Lys Ile Leu Gln Glu Val Thr Ala Gly Gln Ala Val
 195 200 205
 Leu Asn Ile Leu Pro Gly Gly Leu Asp Leu Met Thr Leu Lys Gly Gly
 210 215 220
 Ile Asp Val Ser Arg Val Ala Val Met Gly His Ser Phe Gly Gly Ala
 225 230 235 240
 Thr Ala Ile Leu Ala Leu Ala Lys Glu Met Gln Phe Arg Cys Ala Val
 245 250 255
 Ala Leu Asp Ala Trp Met Phe Pro Leu Glu His Asp Phe Tyr Pro Thr
 260 265 270
 Ala Arg Gly Pro Ile Phe Phe Ile Asn Ala Glu Lys Phe Gln Thr Val
 275 280 285
 Glu Thr Val Asn Leu Met Lys Lys Ile Cys Asp Gln His His Gln Ser
 290 295 300
 Arg Ile Ile Thr Val Leu Gly Ser Val His Arg Ser Leu Thr Asp Phe
 305 310 315 320
 Val Phe Val Ala Gly Asn Trp Ile Ser Lys Phe Phe Ser Ser His Thr
 325 330 335
 Arg Gly Ser Leu Asp Pro Tyr Glu Gly Gln Glu Thr Val Val Arg Ala
 340 345 350
 Met Leu Ala Phe Leu Gln Lys His Leu Asp Leu Lys Glu Asp Tyr Asp
 355 360 365
 Gln Trp Asn Asn Phe Ile Glu Gly Ile Gly Pro Ser Leu Thr Pro Gly
 370 375 380
 Ala Pro His His Leu Ser Ser Leu
 385 390
 (2) INFORMATION FOR SEQ ID NO: 2:
 (i) SEQUENCE CHARACTERISTICS:
 (A) LENGTH: 1665 base pairs
 (B) TYPE: nucleic acid
 (C) STRANDEDNESS: single
 (D) TOPOLOGY: linear
 (ii) MOLECULE TYPE: cDNA
 (vi) ORIGINAL SOURCE:
 (A) ORGANISM: BOVINE (Bos taurus)
 (ix) FEATURE:
 (A) NAME/KEY: CDS
 (B) LOCATION: 111..1286
 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2
 GTCGACCCAC GCGTCCGAGT TGACCGTCTG GGCTGTTTCT GAGGGTCAAC GTGACTCGCC 60
 GTCAAGTTCA GCCACTGCCC AAGTCGTCGT TCAGTTCAGT TGGTTATGAG ATG GGG 116
 Met Gly
 1
 GTC AAC CAG TCT GTG AGC TTC CCA CCC GTC ACG GGA CCC CAC CTC GTA 164
 Val Asn Gln Ser Val Ser Phe Pro Pro Val Thr Gly Pro His Leu Val
 5 10 15
 GGC TGT GGG GAT GTG ATG GAG GGT CAG AGC CTC CAG GGC AGC TTC TTT 212
 Gly Cys Gly Asp Val Met Glu Gly Gln Ser Leu Gln Gly Ser Phe Phe
 20 25 30
 CGA CTG TTC TAC CCG TGC CAA GAG GCA GAG GAG ACC TCG GAG CAG CCC 260
 Arg Leu Phe Tyr Pro Cys Gln Glu Ala Glu Glu Thr Ser Glu Gln Pro
 35 40 45 50
 CTG TGG ATT CCC CGC TAT GAG TAC TGC GCT GGC CTG GCC GAA TAC CTA 308
 Leu Trp Ile Pro Arg Tyr Glu Tyr Cys Ala Gly Leu Ala Glu Tyr Leu
 55 60 65
 AAG TTT AAT AAG CGC TGG GGG GGG TTA CTG TTC AAC CTG GGT GTG GGA 356
 Lys Phe Asn Lys Arg Trp Gly Gly Leu Leu Phe Asn Leu Gly Val Gly
 70 75 80
 TCT TGT CGC CTG CCT GTT AGC TGG AAT GGC CCC TTT AAA ACA AAG GAC 404
 Ser Cys Arg Leu Pro Val Ser Trp Asn Gly Pro Phe Lys Thr Lys Asp
 85 90 95
 TCT GGA TAC CCC TTG ATC ATC TTC TCT CAT GGC ATG GGA GCC TTC AGG 452
 Ser Gly Tyr Pro Leu Ile Ile Phe Ser His Gly Met Gly Ala Phe Arg
 100 105 110
 ACA GTG TAT TCA GCC TTC TGC ATG GAG CTG GCT TCT CGT GGC TTT GTG 500
 Thr Val Tyr Ser Ala Phe Cys Met Glu Leu Ala Ser Arg Gly Phe Val
 115 120 125 130
 GTT GCT GTA CCA GAG CAC AGG GAT GGG TCA GCT GCG GCC ACC TGT TTC 548
 Val Ala Val Pro Glu His Arg Asp Gly Ser Ala Ala Ala Thr Cys Phe
 135 140 145
 TGC AAG CAG ACC CCA GAG GAG AAC CAG CCT GAC AAT GAG GCC CTG AAG 596
 Cys Lys Gln Thr Pro Glu Glu Asn Gln Pro Asp Asn Glu Ala Leu Lys
 150 155 160
 GAG GAA TGG ATC CCC CAC CGT CAA ATT GAG GAA GGG GAG AAG GAA TTC 644
 Glu Glu Trp Ile Pro His Arg Gln Ile Glu Glu Gly Glu Lys Glu Phe
 165 170 175
 TAT GTT CGG AAC TAC CAG GTG CAT CAG AGG GTG AGC GAG TGT GTG AGG 692
 Tyr Val Arg Asn Tyr Gln Val His Gln Arg Val Ser Glu Cys Val Arg
 180 185 190
 GTG TTG AAG ATC CTA CAA GAG GTC ACT GCT GGG CAG GCC GTT CTC AAC 740
 Val Leu Lys Ile Leu Gln Glu Val Thr Ala Gly Gln Ala Val Leu Asn
 195 200 205 210
 ATC TTG CCT GGC GGA TTG GAT CTG ATG ACC TTG AAG GGC GGC ATT GAC 788
 Ile Leu Pro Gly Gly Leu Asp Leu Met Thr Leu Lys Gly Gly Ile Asp
 215 220 225
 GTG AGC CGT GTG GCT GTA ATG GGA CAT TCA TTT GGA GGG GCC ACA GCT 836
 Val Ser Arg Val Ala Val Met Gly His Ser Phe Gly Gly Ala Thr Ala
 230 235 240
 ATT CTG GCC TTG GCC AAG GAG ATG CAA TTT AGG TGT GCT GTG GCT TTG 884
 Ile Leu Ala Leu Ala Lys Glu Met Gln Phe Arg Cys Ala Val Ala Leu
 245 250 255
 GAC GCT TGG ATG TTT CCT CTG GAG CAT GAC TTT TAC CCC ACG GCC CGA 932
 Asp Ala Trp Met Phe Pro Leu Glu His Asp Phe Tyr Pro Thr Ala Arg
 260 265 270
 GGC CCT ATC TTC TTT ATC AAT GCT GAG AAG TTC CAG ACA GTG GAG ACT 980
 Gly Pro Ile Phe Phe Ile Asn Ala Glu Lys Phe Gln Thr Val Glu Thr
 275 280 285 290
 GTC AAC TTG ATG AAA AAG ATT TGT GAC CAG CAC CAC CAA TCC AGG ATC 1028
 Val Asn Leu Met Lys Lys Ile Cys Asp Gln His His Gln Ser Arg Ile
 295 300 305
 ATA ACT GTC CTT GGT TCT GTT CAT CGG AGT CTA ACC GAC TTT GTT TTT 1076
 Ile Thr Val Leu Gly Ser Val His Arg Ser Leu Thr Asp Phe Val Phe
 310 315 320
 GTG GCT GGT AAC TGG ATT AGT AAA TTC TTC TCC AGT CAC ACC CGT GGA 1124
 Val Ala Gly Asn Trp Ile Ser Lys Phe Phe Ser Ser His Thr Arg Gly
 325 330 335
 AGC TTG GAC CCC TAT GAA GGT CAG GAG ACC GTG GTG CGG GCC ATG TTG 1172
 Ser Leu Asp Pro Tyr Glu Gly Gln Glu Thr Val Val Arg Ala Met Leu
 340 345 350
 GCC TTC CTG CAG AAG CAT CTT GAC CTG AAA GAG GAC TAT GAC CAG TGG 1220
 Ala Phe Leu Gln Lys His Leu Asp Leu Lys Glu Asp Tyr Asp Gln Trp
 355 360 365 370
 AAC AAC TTC ATT GAA GGC ATT GGC CCA TCA CTG ACC CCA GGG GCC CCA 1268
 Asn Asn Phe Ile Glu Gly Ile Gly Pro Ser Leu Thr Pro Gly Ala Pro
 375 380 385
 CAC CAT CTG TCC AGC CTG TAGGCACAAC TGGTCATCTT GTGGAAGGTC 1316
 His His Leu Ser Ser Leu
 390
 CCTGAGCTGA GTTCCCGTGT GGGGCCTGCC CAGGGATACC CTTGGCCTCC TATCAGGAAG 1376
 TGATTGCCAT GACCCTTCTG TGTTGATTGA GAGGATATAA TCACACTGCT GATTGGTAAC 1436
 GGGGTACTTG GATTCTCAGA CTTGTCGATC TTAAACTCAT GTTGGGACTT GGGTTCACTT 1496
 ACTGATGGGC AAACGGGCAT TCTGAGGACT GAGCCTTAAT GGTATGGAGA ACAAACAGTG 1556
 GGATGGGGCT GGGGAAGATC TAAGCCCTAA GCTGGGCACT ATGAGCCCTA TAAACCCAAC 1616
 CAGCCAACAC CCTCACCTTG GGCAAGTATG ACTTCTGCAG GTCGACTCT 1665
 (2) INFORMATION FOR SEQ ID NO: 3:
 (i) SEQUENCE CHARACTERISTICS:
 (A) LENGTH: 392 amino acids
 (B) TYPE: amino acid
 (C) STRANDEDNESS: single
 (D) TOPOLOGY: linear
 (ii) MOLECULE TYPE: peptide
 (vi) ORIGINAL SOURCE:
 (A) ORGANISM: HUMAN
 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3
 Met Gly Val Asn Gln Ser Val Gly Phe Pro Pro Val Thr Gly Pro His
 1 5 10 15
 Leu Val Gly Cys Gly Asp Val Met Glu Gly Gln Asn Leu Gln Gly Ser
 20 25 30
 Phe Phe Arg Leu Phe Tyr Pro Cys Gln Lys Ala Glu Glu Thr Met Glu
 35 40 45
 Gln Pro Leu Trp Ile Pro Arg Tyr Glu Tyr Cys Thr Gly Leu Ala Glu
 50 55 60
 Tyr Leu Gln Phe Asn Lys Arg Cys Gly Gly Leu Leu Phe Asn Leu Ala
 65 70 75 80
 Val Gly Ser Cys Arg Leu Pro Val Ser Trp Asn Gly Pro Phe Lys Thr
 85 90 95
 Lys Asp Ser Gly Tyr Pro Leu Ile Ile Phe Ser His Gly Leu Gly Ala
 100 105 110
 Phe Arg Thr Leu Tyr Ser Ala Phe Cys Met Glu Leu Ala Ser Arg Gly
 115 120 125
 Phe Val Val Ala Val Pro Glu His Arg Asp Arg Ser Ala Ala Thr Thr
 130 135 140
 Tyr Phe Cys Lys Gln Ala Pro Glu Glu Asn Gln Pro Thr Asn Glu Ser
 145 150 155 160
 Leu Gln Glu Glu Trp Ile Pro Phe Arg Arg Val Glu Glu Gly Glu Lys
 165 170 175
 Glu Phe His Val Arg Asn Pro Gln Val His Gln Arg Val Ser Glu Cys
 180 185 190
 Leu Arg Val Leu Lys Ile Leu Gln Glu Val Thr Ala Gly Gln Thr Val
 195 200 205
 Phe Asn Ile Leu Pro Gly Gly Leu Asp Leu Met Thr Leu Lys Gly Asn
 210 215 220
 Ile Asp Met Ser Arg Val Ala Val Met Gly His Ser Phe Gly Gly Ala
 225 230 235 240
 Thr Ala Ile Leu Ala Leu Ala Lys Glu Thr Gln Phe Arg Cys Ala Val
 245 250 255
 Ala Leu Asp Ala Trp Met Phe Pro Leu Glu Arg Asp Phe Tyr Pro Lys
 260 265 270
 Ala Arg Gly Pro Val Phe Phe Ile Asn Thr Glu Lys Phe Gln Thr Met
 275 280 285
 Glu Ser Val Asn Leu Met Lys Lys Ile Cys Ala Gln His Glu Gln Ser
 290 295 300
 Arg Ile Ile Thr Val Leu Gly Ser Val His Arg Ser Gln Thr Asp Phe
 305 310 315 320
 Ala Phe Val Thr Gly Asn Leu Ile Gly Lys Phe Phe Ser Thr Glu Thr
 325 330 335
 Arg Gly Ser Leu Asp Pro Tyr Glu Gly Gln Glu Val Met Val Arg Ala
 340 345 350
 Met Leu Ala Phe Leu Gln Lys His Leu Asp Leu Lys Glu Asp Tyr Asn
 355 360 365
 Gln Trp Asn Asn Leu Ile Glu Gly Ile Gly Pro Ser Leu Thr Pro Gly
 370 375 380
 Ala Pro His His Leu Ser Ser Leu
 385 390
 (2) INFORMATION FOR SEQ ID NO: 4:
 (i) SEQUENCE CHARACTERISTICS:
 (A) LENGTH: 2559 base pairs
 (B) TYPE: nucleic acid
 (C) STRANDEDNESS: single
 (D) TOPOLOGY: linear
 (ii) MOLECULE TYPE: cDNA
 (vi) ORIGINAL SOURCE:
 (A) ORGANISM: HUMAN
 (ix) FEATURE:
 (A) NAME/KEY: CDS
 (B) LOCATION: 216..1392
 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4
 GCAGGTCTCG ACCCACGCGT CCGCGGACGC GTGGGCGAGA AGTGCTTCCA AGCGTCCATT 60
 TTGAGCCTTG GAAACTACGA CGACCAAAGG GCCACGGGTT CCTGGGTCGT TTCTCATTTC 120
 CGTCGAGTTA AACGTCTGGG GCTGCTTCTG AGGAATCAGC TTGGCTGGCC AGCAAGTTCA 180
 GCTCCGGCAA GTCATTTGAT TCACCCGGTG ATGAA ATG GGG GTC AAC CAG TCT 233
 Met Gly Val Asn Gln Ser
 1 5
 GTG GGC TTT CCA CCT GTC ACA GGA CCC CAC CTC GTA GGC TGT GGG GAT 281
 Val Gly Phe Pro Pro Val Thr Gly Pro His Leu Val Gly Cys Gly Asp
 10 15 20
 GTG ATG GAG GGT CAG AAT CTC CAG GGG AGC TTC TTT CGA CTC TTC TAC 329
 Val Met Glu Gly Gln Asn Leu Gln Gly Ser Phe Phe Arg Leu Phe Tyr
 25 30 35
 CCC TGC CAA AAG GCA GAG GAG ACC ATG GAG CAG CCC CTG TGG ATT CCC 377
 Pro Cys Gln Lys Ala Glu Glu Thr Met Glu Gln Pro Leu Trp Ile Pro
 40 45 50
 CGC TAT GAG TAC TGC ACT GGC CTG GCC GAG TAC CTG CAG TTT AAT AAG 425
 Arg Tyr Glu Tyr Cys Thr Gly Leu Ala Glu Tyr Leu Gln Phe Asn Lys
 55 60 65 70
 CGC TGC GGG GGC TTG CTG TTC AAC CTG GCG GTG GGA TCT TGT CGC CTG 473
 Arg Cys Gly Gly Leu Leu Phe Asn Leu Ala Val Gly Ser Cys Arg Leu
 75 80 85
 CCT GTT AGC TGG AAT GGC CCC TTT AAG ACA AAG GAC TCT GGA TAC CCC 521
 Pro Val Ser Trp Asn Gly Pro Phe Lys Thr Lys Asp Ser Gly Tyr Pro
 90 95 100
 TTG ATC ATC TTC TCC CAT GGC CTA GGA GCC TTC AGG ACT TTG TAT TCA 569
 Leu Ile Ile Phe Ser His Gly Leu Gly Ala Phe Arg Thr Leu Tyr Ser
 105 110 115
 GCC TTC TGC ATG GAG CTG GCC TCA CGT GGC TTT GTG GTT GCT GTG CCA 617
 Ala Phe Cys Met Glu Leu Ala Ser Arg Gly Phe Val Val Ala Val Pro
 120 125 130
 GAG CAC AGG GAC CGG TCA GCG GCA ACC ACC TAT TTC TGC AAG CAG GCC 665
 Glu His Arg Asp Arg Ser Ala Ala Thr Thr Tyr Phe Cys Lys Gln Ala
 135 140 145 150
 CCA GAA GAG AAC CAG CCC ACC AAT GAA TCG CTG CAG GAG GAA TGG ATC 713
 Pro Glu Glu Asn Gln Pro Thr Asn Glu Ser Leu Gln Glu Glu Trp Ile
 155 160 165
 CCT TTC CGT CGA GTT GAG GAA GGG GAG AAG GAA TTT CAT GTT CGG AAT 761
 Pro Phe Arg Arg Val Glu Glu Gly Glu Lys Glu Phe His Val Arg Asn
 170 175 180
 CCC CAG GTG CAT CAG CGG GTA AGC GAG TGT TTA CGG GTG TTG AAG ATC 809
 Pro Gln Val His Gln Arg Val Ser Glu Cys Leu Arg Val Leu Lys Ile
 185 190 195
 CTG CAA GAG GTC ACT GCT GGG CAG ACT GTC TTC AAC ATC TTG CCT GGT 857
 Leu Gln Glu Val Thr Ala Gly Gln Thr Val Phe Asn Ile Leu Pro Gly
 200 205 210
 GGC TTG GAT CTG ATG ACT TTG AAG GGC AAC ATT GAC ATG AGC CGT GTG 905
 Gly Leu Asp Leu Met Thr Leu Lys Gly Asn Ile Asp Met Ser Arg Val
 215 220 225 230
 GCT GTG ATG GGA CAT TCA TTT GGA GGG GCC ACA GCT ATT CTG GCT TTG 953
 Ala Val Met Gly His Ser Phe Gly Gly Ala Thr Ala Ile Leu Ala Leu
 235 240 245
 GCC AAG GAG ACC CAA TTT CGG TGT GCG GTG GCT CTG GAT GCT TGG ATG 1001
 Ala Lys Glu Thr Gln Phe Arg Cys Ala Val Ala Leu Asp Ala Trp Met
 250 255 260
 TTT CCT CTG GAA CGT GAC TTT TAC CCC AAG GCC CGA GGA CCT GTG TTC 1049
 Phe Pro Leu Glu Arg Asp Phe Tyr Pro Lys Ala Arg Gly Pro Val Phe
 265 270 275
 TTT ATC AAT ACT GAG AAA TTC CAG ACA ATG GAG AGT GTC AAT TTG ATG 1097
 Phe Ile Asn Thr Glu Lys Phe Gln Thr Met Glu Ser Val Asn Leu Met
 280 285 290
 AAG AAG ATA TGT GCC CAG CAT GAA CAG TCT AGG ATC ATA ACC GTT CTT 1145
 Lys Lys Ile Cys Ala Gln His Glu Gln Ser Arg Ile Ile Thr Val Leu
 295 300 305 310
 GGT TCT GTT CAT CGG AGT CAA ACT GAC TTT GCT TTT GTG ACT GGC AAC 1193
 Gly Ser Val His Arg Ser Gln Thr Asp Phe Ala Phe Val Thr Gly Asn
 315 320 325
 TTG ATT GGT AAA TTC TTC TCC ACT GAA ACC CGT GGG AGC CTG GAC CCC 1241
 Leu Ile Gly Lys Phe Phe Ser Thr Glu Thr Arg Gly Ser Leu Asp Pro
 330 335 340
 TAT GAA GGG CAG GAG GTT ATG GTA CGG GCC ATG TTG GCC TTC CTG CAG 1289
 Tyr Glu Gly Gln Glu Val Met Val Arg Ala Met Leu Ala Phe Leu Gln
 345 350 355
 AAG CAC CTC GAC CTG AAA GAA GAC TAT AAT CAA TGG AAC AAC CTT ATT 1337
 Lys His Leu Asp Leu Lys Glu Asp Tyr Asn Gln Trp Asn Asn Leu Ile
 360 365 370
 GAA GGC ATT GGA CCG TCG CTC ACC CCA GGG GCC CCC CAC CAT CTG TCC 1385
 Glu Gly Ile Gly Pro Ser Leu Thr Pro Gly Ala Pro His His Leu Ser
 375 380 385 390
 AGC CTG T AGGCACAACT GGCCATTTGT AAAGTCACTT CAGCCAAGTT TTCATTTGGG 1442
 Ser Leu
 AGCTACCCAA GGGCACCCAT GAGCTCCTAT CAAGAAGTGA TCAACGTGAC CCCTTTTCAC 1502
 AGATTGAAAG GTGTAATCAC ACTGCTGCTT GGATAACTGG GTACTTTGAT CTTAGATTTG 1562
 ATCTTAAAAT CACTTTGGGA CTGGGATCCC TTGCTGATTG ACAAACAGAC TTTCTGGGAC 1622
 CTTGATGGAG TGGGGAACAA GCAGTAGAGT GGGACTGGGG GAGACCCAGG CCCCGGGCTG 1682
 AGCACTGTGA GGCCTGGATG TGAAGACTCA GCCCAGCGAA GCTCATTCCC TTACCCCCGG 1742
 CCAGTGCTGC TGCTTCAGTG GAAGAGATGA AGCCAAAGGA CAGAATGAAA ATCCCTACCT 1802
 TCAGAGACTC TAGCCCAGCC CAACACCATC TCTTCCTACC TCTCAGCCTT CTCCCTCCCC 1862
 AGGGCCACTT GTTGAAGTCT GAGCACTTTA TGTAAATTTC TAGGTGTGAG CCGTGATCAC 1922
 ATTTTCTATT TATTTCCAAG TCTTCTCATT GTATGGAACA TAGTACTACT TATACTTACA 1982
 GTAGTAAGTT ATACTTGTGA GCCCACAGAG TGGCAGACAG CATGGCTCTC ACAGCACAGG 2042
 GAGAAAAACT GAGGTACACA GAGGTACCTC AGAAGCTCTG GATGTCTTTG GGGGTTTTGC 2102
 TAAGTGTATC TTGATAGGAA ACAACAAAAG CAGGTTGAGA TGGGGAAGAT GACAGAACAA 2162
 CAGTGTTAAA TGGCCATTTG CACAGGCCTT TGCCACAACA GAGAAGTAGT TTGGTCAGCT 2222
 AAAACTCAGC TGCAGCCTGG ACAGTAGAGC GAGACCCCAT CTTAAAAATA AAGAAGGCTG 2282
 GGCGTGGTGG CTCATGCCTG TAATCCCAGC ACTTTGGGAG GCCAAGGCAG GCAGATCACT 2342
 TAAGGCCAGG AGTTCAAGAC CACCTGGCCA ACATGGTGAA ACCCCGTCTC TACTAAAAAT 2402
 ACAAAAAATT AGCCTGGCGT AATGGCAGGC GCCTATAATC CCAGCTACTC AGGAGGCTGA 2462
 AGCAGAAGAA TCACTTGAAC CTAGGAGGCG GAGGTTGCAG TGAGTCAAGA TCGCGCCACT 2522
 GCACTCCAGC CTGGGTGACA GAGCAAGACT CTGTCTT 2559