Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/36297
標題: 利用桿狀病毒/Sf21昆蟲細胞表現系統功能性表現與特性分析黑腹胡蜂之二肽基胜肽酶
Functional expression and characterization of dipeptidyl peptidase IV from the black-bellied hornet Vespa basalis in Sf21 Insect Cells
作者: 謝聖國
Hsieh, Sheng-Kuo
關鍵字: dipeptidyl peptidase IV
二肽基胜肽酶蜂毒肽轉譯後修飾
mastoparan B
prosequence processing
sitagliptin
Vespa basalis
西格列汀
黑腹胡蜂
出版社: 生物科技學研究所
引用: 和七一, 余曉東. 2007. 胡蜂蜂毒肽B的研究概况. Journal of Chongqing Normal University (Natural Science Edition). 24:1-5. 靳子蓉, 高穗生. 桿狀病毒表現載體之發展與應用. 行政院農業委員會農業藥物毒物試驗所技術專刊. 第137 號. Abbott, C.A., E. Baker, G.R. Sutherland, and G.W. McCaughan. 1994. Genomic organization, exact localization, and tissue expression of the human CD26 (dipeptidyl peptidase IV) gene. Immunogenetics. 40:331-338. Aertgeerts, K. 2004. N-linked glycosylation of dipeptidyl peptidase IV (CD26): Effects on enzyme activity, homodimer formation, and adenosine deaminase binding. Protein Science. 13:145-154. Aertgeerts, K., S. Ye, L. Shi, S.G. Prasad, D. Witmer, E. Chi, B.C. Sang, R.A. Wijnands, D.R. Webb, and R.V. Swanson. 2004. N-linked glycosylation of dipeptidyl peptidase IV (CD26): effects on enzyme activity, homodimer formation, and adenosine deaminase binding. Protein Sci. 13:145-154. Ayres, M.D., S.C. Howard, J. Kuzio, M. Lopez-Ferber, and R.D. Possee. 1994. The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology. 202:586-605. Barrett, A.J., and N.D. Rawlings. 1992. Oligopeptidases, and the emergence of the prolyl oligopeptidase family. Biol Chem Hoppe Seyler. 373:353-360. C.H. Ahrens, D.J. Leisy, and G.F. Rohrmann. 1996. DNA Replication in Eukaryotic Cells Chapter 31 Baculovirus DNA Replication. Cold Spring Harbor Laboratory Press 855-872. Cha, H.J., N.G. Dalal, V.N. Vakharia, and W.E. Bentley. 1999. Expression and purification of human interleukin-2 simplified as a fusion with green fluorescent protein in suspended Sf-9 insect cells. J Biotechnol. 69:9-17. Chahdi, A., W.S. Choi, Y.M. Kim, and M.A. Beaven. 2003. Mastoparan selectively activates phospholipase D2 in cell membranes. J Biol Chem. 278:12039-12045. De Meester, I., S. Korom, J. Van Damme, and S. Scharpe. 1999. CD26: Let it cut or cut it down. immunol Today. 20:367-375. Dooseop Kim, Liping Wang, Maria Beconi, George J. Eiermann, Michael H. Fisher, Huaibing He, Gerard J. Hickey, Jennifer E. Kowalchick, Barbara Leiting, Kathryn Lyons, Frank Marsilio, Margaret E. McCann, Reshma A. Patel, Aleksandr Petrov, Giovanna Scapin, Sangita B. Patel, Ranabir Sinha Roy, Joseph K. Wu, Matthew J. Wyvratt, Bei B. Zhang, Lan Zhu, Nancy A. Thornberry, and A.E. Weber. 2005. (2R)-4-Oxo-4-[3-(Trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine: A Potent, Orally Active Dipeptidyl Peptidase IV Inhibitor for the Treatment of Type 2 Diabetes. J. Med. Chem. 48:141-151. Eipper, B.A., S.L. Milgram, E.J. Husten, H.Y. Yun, and R.E. Mains. 1993. Peptidylglycine alpha-amidating monooxygenase: a multifunctional protein with catalytic, processing, and routing domains. Protein Sci. 2:489-497. Engel, M., T. Hoffmann, L. Wagner, M. Wermann, U. Heiser, R. Kiefersauer, R. Huber, W. Bode, H.U. Demuth, and H. Brandstetter. 2003. The crystal structure of dipeptidyl peptidase IV (CD26) reveals its functional regulation and enzymatic mechanism. Proc Natl Acad Sci U S A. 100:5063-5068. Gorrell, M.D. 2005. Dipeptidyl peptidase IV and related enzymes in cell biology and liver disorders. Clin Sci (Lond). 108:277-292. Higashijima, T., J. Burnier, and E.M. Ross. 1990. Regulation of Gi and Go by mastoparan, related amphiphilic peptides, and hydrophobic amines. Mechanism and structural determinants of activity. J Biol Chem. 265:14176-14186. Hiramatsu, H., K. Kyono, H. Shima, C. Fukushima, S. Sugiyama, K. Inaka, A. Yamamoto, and R. Shimizu. 2003. Crystallization and preliminary X-ray study of human dipeptidyl peptidase IV (DPPIV). Acta Crystallogr D Biol Crystallogr. 59:595-596. Ho, C.-L., Y.-L. LIN, W.-C. CHEN, H.-M. YU, K.-T. WANG, L.-L. HWANG, and C.-T. CHEN. 1995. Immunogenicity of Mastoparan B, a Cationic Tetradecapeptide Isolated From the Hornet (Vespa basalis) Venom, and its Structural Requirements. Toxicon. 33:1443-1451. Ho, C.L., and L.L. Hwang. 1991. Structure and biological activities of a new mastoparan isolated from the venom of the hornet Vespa basalis. Biochem J. 274 ( Pt 2):453-456. Ho, C.L., L.L. Hwang, Y.L. Lin, C.T. Chen, H.M. Yu, and K.T. Wang. 1994. Cardiovascular effects of mastoparan B and its structural requirements. Eur J Pharmacol. 259:259-264. Ho, C.L., Y.L. LIN, W.C. CHEN, L.L. HWANG, H.M. YU, and K.T. WANG. 1996. Structural requirements for the edema-inducing and hemolytic activities of mastoparan B isolated form the hornet (Vespa basalis) venom. Toxicon. 34:1027-1035. Ho, C.L., Y.P. Shih, K.T. Wang, and H.M. Yu. 2001. Enhancing the hypotensive effect and diminishing the cytolytic activity of hornet mastoparan B by D-amino acid substitution. Toxicon. 39:1561-1566. Hofmann, C., V. Sandig, G. Jennings, M. Rudolph, P. Schlag, and M. Strauss. 1995. Efficient gene transfer into human hepatocytes by baculovirus vectors. Proc Natl Acad Sci U S A. 92:10099-10103. Hopsu-Havu, V.K., and G.G. Glenner. 1966. A new dipeptide naphthylamidase hydrolyzing glycyl-prolyl-beta-naphthylamide. Histochemie. 7:197-201. Jang, S., T.Y. Chung, J. Shin, K.L. Lin, J.T. Tzen, and F.Y. Li. 2010. Docking study of the precursor peptide of mastoparan onto its putative processing enzyme, dipeptidyl peptidase IV: a revisit to molecular ticketing. J Comput Aided Mol Des. 24:213-224. Kabashima, T., K. Ito, and T. Yoshimoto. 1996. Dipeptidyl Peptidase IV from Xanthomonas maltophilia: Sequencing and Expression of the Enzyme Gene and Characterization of the Expressed Enzyme. J. Biochem. 120:1111-1117. Kabashima, T., T. Yoshida, K. Ito, and T. Yoshimoto. 1995. Cloning, Sequencing, and Expression of the Dipeptidyl Peptidase IV Gene from Flavoacterium meningosepticu in Escherichia coli. Archives of Biochemistry and Biophysics. 320:123-128. Kikkawa, F., H. Kajiyama, K. Shibata, K. Ino, S. Nomura, and S. Mizutani. 2005a. Dipeptidyl peptidase IV in tumor progression. Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics. 1751:45-51. Kikkawa, F., H. Kajiyama, K. Shibata, K. Ino, S. Nomura, and S. Mizutani. 2005b. Dipeptidyl peptidase IV in tumor progression. Biochim Biophys Acta. 1751:45-51. Kreil, G., L. Haiml, and G. Suchanek. 1980. Stepwise cleavage of the pro part of promelittin by dipeptidylpeptidase IV. Evidence for a new type of precursor--product conversion. Eur J Biochem. 111:49-58. Kreil, G., and G. Kreil-Kiss. 1967. The isolation of N-formylglycine from a polypeptide present in bee venom. Biochem Biophys Res Commun. 27:275-280. Langer, J., and S. Ansorge. 2002. Ectopeptidases: CD13/ Aminopeptidase N and CD26/DipeptidylpeptidaseIV in medicine and biology. Kluwer/Plenum, New York. Lee, V.S.Y., W.C. Tu, T.R. Jinn, C.C. Peng, L.J. Lin, and J.T.C. Tzen. 2007. Molecular cloning of the precursor polypeptide of mastoparan B and its putative processing enzyme, dipeptidyl peptidase IV, from the black-bellied hornet, Vespa basalis. Insect Molecular Biology. 16:231-237. Maeda, S. 1989. Expression of foreign genes in insects using baculovirus vectors. Annu Rev Entomol. 34:351-372. McDonald, J.K., and A.J. Barrett. 1986. Mammalian Proteases:A Glossary and Bibliography, Vol. 2, Exopeptidases. Academic Press, London.:111-144. Merkler, D.J. 1994. C-terminal amidated peptides: production by the in vitro enzymatic amidation of glycine-extended peptides and the importance of the amide to bioactivity. Enzyme Microb Technol. 16:450-456. Mollay, C., U. Vilas, A. Hutticher, and G. Kreil. 1986. Isolation of a dipeptidyl aminopeptidase, a putative processing enzyme, from skin secretion of Xenopus laevis. Eur J Biochem. 160:31-35. Murphy, C.I., and H. Piwnica-Worms. 1995. Overview of the Baculovirus UNIT 5.4 Expression System. Current Protocols in Protein Science:5.4.1-5.4.4. Nemoto, E., S. Sugawara, H. Takada, S. Shoji, and H. Horiuch. 1999. Increase of CD26/dipeptidyl peptidase IV expression on human gingival fibroblasts upon stimulation with cytokines and bacterial components. Infect Immun. 67:6225-6233. Neumiller, J.J. 2009. Differential chemistry (structure), mechanism of action, and pharmacology of GLP-1 receptor agonists and DPP-4 inhibitors. Journal of the American Pharmacists Association. 49:S16-S29. O’Reilly, D.R., L.K. Miller, and V.A. luckow. 1992. Baculovirus expression vector: A laboratory manual. . W.H. freeman and company Press. New York. Oravecz, T., M. Pall, G. Roderiquez, M.D. Gorrell, M. Ditto, N.Y. Nguyen, R. Boykins, E. Unsworth, and M.A. Norcross. 1997. Regulation of the receptor specificity and function of the chemokine RANTES (regulated on activation, normal T cell expressed and secreted) by dipeptidyl peptidase IV (CD26)-mediated cleavage. J Exp Med. 186:1865-1872. Park, N.G., Y. Yamato, S. Lee, and G. Sugihara. 1995. Interaction of mastoparan-B from venom of a hornet in Taiwan with phospholipid bilayers and its antimicrobial activity. Biopolymers. 36:793-801. Pascual, I., H. Gomez, T. Pons, M. Chappe, M.A. Vargas, G. Valdes, A. Lopez, A. Saroyan, J.L. Charli, and M. de los Angeles Chavez. 2011. Effect of divalent cations on the porcine kidney cortex membrane-bound form of dipeptidyl peptidase IV. Int J Biochem Cell Biol. 43:363-371. Piek, T., and P. Mantel. 1986. Cholinergic antagonists in a solitary wasp venom. Comp Biochem Physiol C. 85:433-436. Raz, I., M. Hanefeld, L. Xu, C. Caria, D. Williams-Herman, and H. Khatami. 2006. Efficacy and safety of the dipeptidyl peptidase-4 inhibitor sitagliptin as monotherapy in patients with type 2 diabetes mellitus. Diabetologia. 49:2564-2571. Rea, D., and V. Fulop. 2006. Structure–Function Properties of Prolyl Oligopeptidase Family Enzymes Cell Biochemistry and Biophysics. 44:349-365. Scheen, A.J. 2010. Pharmacokinetics of dipeptidylpeptidase-4 inhibitors. Diabetes, Obesity and Metabolism. 12:648-658. Schmidt, J.O. 1982. Biochemistry of insect venoms. Annu Rev Entomol. 27:339-368. Scott, R., M. Wu, M. Sanchez, and P. Stein. 2007. Efficacy and tolerability of the dipeptidyl peptidase-4 inhibitor sitagliptin as monotherapy over 12 weeks in patients with type 2 diabetes. Int J Clin Pract. 61:171-180. Sehgal, D., P.S. Malik, and S. Jameel. 2003. Purification and diagnostic utility of a recombinant hepatitis E virus capsid protein expressed in insect larvae. Protein Expr Purif. 27:27-34. Smith, G.E., M.D. Summers, and M.J. Fraser. 1983. Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol Cell Biol. 3:2156-2165. Suchanek, G., G. Kreil, and M.A. Hermodson. 1978. Amino acid sequence of honeybee prepromelittin synthesized in vitro. Proc Natl Acad Sci U S A. 75:701-704. Sukumar, M., and T. Higashijima. 1992. G protein-bound conformation of mastoparan-X, a receptor-mimetic peptide. J Biol Chem. 267:21421-21424. Tsakalidou, E., R. Anastasiou, K. Papadimitriou, E. Manolopoulou, and G. Kalantzopoulos. 1997. Purification and characterisation of an intracellular X-prolyl-dipeptidyl aminopeptidase from Streptococcus thermophilus ACA-DC 4. J Biotechnol. 59:203-211. Tuan, S.J., S.S. Kao, and D.J. Cheng. 1994. Histopathology and pathogenicity of Spodoptera wxigua nuclear polyhedrosis virus isolated in Taiwan. Chinese J. Entomol. 14:33-45. White, J.R., Jr, PA, and PharmD. 2008. Dipeptidyl Peptidase-IV Inhibitors: Pharmacological Profile and Clinical Use. Clinical Diabetes. 26:53-57. Whitford, M., S. Stewart, J. Kuzio, and P. Faulkner. 1989. Identification and sequence analysis of a gene encoding gp67, an abundant envelope glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol. 63:1393-1399. Wiley, J., and L. Sons. 2009. Dipeptidyl peptidase-4 (DPP-4) inhibitors for type 2 diabetes mellitus The Cochrane Collaboration:1-154. Yazbeck, R., G.S. Howarth, and C.A. Abbott. 2009. Dipeptidyl peptidase inhibitors, an emerging drug class for inflammatory disease? Trends in Pharmacological Sciences. 30:600-607. Ye, Y., K.T. Keyes, C. Zhang, J.R. Perez-Polo, Y. Lin, and Y. Birnbaum. 2010. The myocardial infarct size-limiting effect of sitagliptin is PKA-dependent, whereas the protective effect of pioglitazone is partially dependent on PKA. Am J Physiol Heart Circ Physiol. 298:H1454-1465. Yoshimoto, T., and D. Tsuru. 1982. Proline-specific dipeptidyl aminopeptidase from Flavobacterium meningosepticum. J Biochem. 91:1899-1906. Yu, D.M.T., T.-W. Yao, S. Chowdhury, N.A. Nadvi, B. Osborne, W.B. Church, G.W. McCaughan, and M.D. Gorrell. 2010. The dipeptidyl peptidase IV family in cancer and cell biology. FEBS Journal. 277:1126-1144. Zhang, Y., P. McAtee, P.O. Yarbough, A.W. Tam, and T. Fuerst. 1997. Expression, characterization, and immunoreactivities of a soluble hepatitis E virus putative capsid protein species expressed in insect cells. Clin Diagn Lab Immunol. 4:423-428. Zhou, N., Y. Zhang, W. Jing, Z. Li, and X. Wu. 1995. High expression of HBV S gene in Bombyx mori cell culture and in silkworms. Chin J Biotechnol. 11:149-156.
摘要: Mastoparan B是黑腹胡蜂(black-bellied hornet; Vespa basalis)毒素中最主要的成分,其prosequnce推測是經由二肽基胜肽酶(Dipeptidyl peptidase IV; DPP-IV)的水解作用進行轉譯後的修飾。而此二肽基胜肽酶的互補DNA已於2007年由本實驗室成功的從黑腹胡蜂的腺體中分離及選殖。該黑腹胡蜂二肽基胜肽酶的蛋白質序列經分析發現和人類的二肽基胜肽酶的三級結構極為相似。 此篇論文乃是利用桿狀病毒/昆蟲細胞表現系統表現來自黑腹胡蜂的二肽基胜肽酶。此重組的二肽基胜肽酶(rDPP-IV)藉用重組的桿狀病毒株vAcP10DPPIV於Sf21昆蟲細胞表現並以鎳親合性管柱層析純化,再經由西方墨點法和LC-MS/MS確認。該rDPP-IV在m.o.i.=10感染96小時有最佳的活性表現。再經由鎳親合性管柱層析純化rDPP-IV,並測得增加五倍的比活性。針對五種不同的反應物測得其kcat/Km的範圍從10-500 mM-1.S-1。該二肽基胜肽酶rDPP-IV最佳活性表現的溫度與pH分別為50℃和pH 9。而sitagliptin(DPP-IV抑制劑)和phenylmethylsulfonyl fluoride(絲氨酸蛋白酶抑制劑)對於二肽基胜肽酶酵素活性分別抑制了80%和60%。經由本研究可以確認所選殖的二肽基胜肽酶是具有類似此serine protease的酵素活性。
Maturation of mastoparan B, the major toxin peptide in the venom of Vespa basalis, requires an enzymatic cleavage of its prosequence presumably via sequential liberation of dipeptides. A putative dipeptidyl peptidase IV cDNA fragment was cloned from Vespa basalis (Black-bellied hornet) venom gland in 2007. From sequence comparison reveals that the DPP-IV from Vespa basalis is homologous to a human DPP-IV whose three-dimensional structure has been determined by X-ray crystallography. In this study, dipeptidyl peptidase IV, was expressed as a glycosylated His-tag fusion protein (rDPP-IV) via the baculovirus expression system. The rDPP-IV purified by one-step nickel-affinity chromatography was verified by Western blot and LC-MS/MS analysis. When Sf21 insect cell was infected by vAcP10DPPIV m.o.i.=10, 96 hours is the optimal for enzyme activity. rDPP-IV specific activity approached 5 times when be purified by nickel-affinity chromatography. The kcat/Km of rDPP-IV was determined to be in the range of 10-500 mM-1.S-1 for five synthetic substrates. The optimal temperature and pH for rDPP-IV were determined to be 50℃ and pH 9. Enzymatic activity of rDPP-IV was significantly reduced by 80% and 60% in the presence of sitagliptin and phenylmethylsulfonyl fluoride, respectively. According to this study, the specific amino acid sequences can digestion by rDPP-IV.
URI: http://hdl.handle.net/11455/36297
其他識別: U0005-2206201118234300
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