Grant-in-Aid for International Scientific Research.
|Section||Joint Research .|
|Research Institution||Tokyo University of Pharmacy and Life Science|
TAKAHASHI Kenji Tokyo University of Pharmacy and Life Science, School of Life Science, Professor, 生命科学部, 教授 (70011533)
DISSANAYAKE ハーバード大学, 公衆衛生学部, 研究員
WIMALASIRI W ペラデニア大学, 医学部, 講師
ATHAUDA Sena ペラデニア大学, 医学部, 講師
KOJIMA Masaki Tokyo University of Pharmacy and Life Science, School of Life Science, Assistant, 生命科学部, 助手 (90277252)
INOUE Hideshi Tokyo University of Pharmacy and Life Science, School of Life Science, Assistant, 生命科学部, 助教授 (20184765)
SENARATH Dis ハーバード大学, 公衆衛生学部, 研究員
WEELAPPULI R ペラデニア大学, 医学部, 講師
SENARATH B.P ペラデニア大学, 医学部, 講師
SENARCTH Dis ハーバード大学, 公衆衛生学部, 研究員
R.WEERAPPULI ペラデニア大学, 医学部, 講師
ATHOUDA Senarath B.P. University of Peradeniya, Department of Medicine, Lecturer
WINALASIRI Weerappuli R. University of Peradeniya, Department of Medicine, Lecturer
DISSONAYAKE Senarath Harvard University, Department of Public Health, Research Associate
|Project Fiscal Year
1996 – 1998
Completed(Fiscal Year 1998)
|Budget Amount *help
¥8,000,000 (Direct Cost : ¥8,000,000)
Fiscal Year 1998 : ¥2,300,000 (Direct Cost : ¥2,300,000)
Fiscal Year 1997 : ¥2,700,000 (Direct Cost : ¥2,700,000)
Fiscal Year 1996 : ¥3,000,000 (Direct Cost : ¥3,000,000)
|Keywords||Filarial proteinase / cDNA Library / Aspartic proteinase / Cryteine proteinase / Cloning / Base sequence analysis / Human filaria / Bovine filaria / フィラリア・プロテアーゼ / cDNAライブラリー / アスパラギン酸プロテアーゼ / システイン・プロテアーゼ / クローニング / 塩基配列決定 / ヒト・フィラリア / ウシ・フィラリア / フィラリアプロテアーゼ / Brugia malayi / システインプロテアーゼ / カテプシンロ / ガストリックシン / カテプシンS / カテプシンD / 塩基配列解析 / フィラリア / プロテアーゼ / PCR|
1.Two aspartic proteinases were purified from the crude extract of boviae filarial parasites (Setcria digitata) collected in Sri Lanka by sucessive steps of chromatography on columns of DEAE-cellulose, Sephacryl S-200, pepstatia-Sepharose, and monoQ, and their enzymat.ic and molecular properties were investigated and the N-terminal sequences were determined. In addition, neutral proteinases were partially purified.
2.cDNA libraries were prepared from the mRNAs of human filarial parasites, including Brugia malayi. Ozwhocerca volvulus and Brugia phangi, and genomic DNA libraries from Brugia malayi and Wuchereria bancrofti. Then, full-length cDNAs of two aspartic proteinases, Asp2 and Asp3. and a partial length cDNA of one aspartic proteinase, Aspl, were isolated from the cDNA libraries of Brugia malayi and their base sequences were determined and amin acid sequences deduced. A phylogenetic tree of aspartic proteinases includingAsp2 and Asp3 was constructed. indicating. that Asp2 somewhat
resembles mammalian cathepsin D, whereas Asp3 is unique in that it is significantly diverged from both cathepsin D and pepsin. AspI was shown to be fairly similar in sequence to Asp2.
3, From the cDNA library of Brugia malayi, full-length cDNAs of two cysteine proteinases, Cysl and Cys2, were isolated, and their base sequences were determined and amino acid sequences deduced. The sequences are fairly similar between Cys1 and Cys2, but are markedly different from those of mammalian cysteine proteinases.
4.In addition, purification and/or characterization studies were performed for a novel inembrane-bound inetalloendopeptidase cleaving progastrin C-terminal (88-101) peptide from bovine brain, a porcine brain endopeptidase specific for the C-terminal CAAX motif of Ras and related proteins, porcine . enteropeptidase, pancreatic trypsin and alpha-chymotrypsin, a rat liver microsomal membrane- bound serine proteinase(hepsin), colopsin (a novel membrane-bound serine proteinase from intestine), human and mouse cathepsin E, coelacant pepsinogen, nope nthesin, mold acid proteinases A and B, bacterial metalloendopeptidases, sigaital peptidase and brotnelain inhibitors. Less