Epitope domain and plasminogen activating site (s) in streptokinase molecule

链激酶分子中的表位结构域和纤溶酶原激活位点

• 批准号: 05670268
• 负责人: OHKUNI Hisashi
• 金额: $ 1.15万
• 依托单位: Nippon Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670268/
• 关键词: straptokinase; epitope; acute glomerulonephritis; plasminogen

We have previously reported that a monoclonal antibody (mAb), N-59, prepared against streptokinase (SKase) from a group A,M type 12 streptococcal isolate (strain A374) from a patient with poststreptococcal glomerulonephritis (PSGN) recognized an epitope also present in SKase from a group C Streptococcus (strain H46A) which is not a PSGN isolate.However, another mAb, RU-1, prepared against SKase (nephritis strain-assiciated SKase, NSA-SKase) from strain A374 reacted only with the homologous SKase and not with common-SKase (C-SKase) from strain H46A.In addition, the complete amino acid sequence of the NSA-SKase of strain A374 was determined, and the epitope domain specific for RU-1 mAb reacted only with the NSA-SKase was localized to residues 164-236.Both pin technology and ELISA test were used to the determination of epitope domain involved in the internal region of NSA-SKase molecule reacted with RU-1 mAb. As the results, RU-1 mAb reacted with residues 177-185 (PSLKERYHL), and the minimum epitope domain, which reacts with RU-1 mAb, was residues 177-180 (PSLK).The other side, N-59 mAb reacted with almost all of SKase (C-SKase). The epitope domain of C-SKase reacted with N-59 mAb was analyzed. As the results, N-59 mAb recognized residues 324-332 (RDLYDPRDK) in C-SKase molecule, and the minimum epitope domain was residues 324-331 (RDLYDPRD).

我们曾报道过一株来自A、M-12型链球菌分离株(A374株)的抗链球菌酶(Skase)的单抗N-59，该单抗可识别非PSGN分离株的C群链球菌(H46A株)的Skase表位。而从A374株制备的抗Skase(肾炎株相关的Skase，NSA-Skase)的单抗RU-1仅与同源Skase反应，而不与H46A株的普通Skase(C-Skase)反应测定了A374株NSA-Skase的全长氨基酸序列，并将RU-1mAb仅与NSA-Skase反应的表位结构域定位在164-236位。结果表明，RU-1单抗与177-185位残基(PSLKERYHL)反应，最小表位区域为177-180位(PSLK)，N-59位单抗与Skase几乎全部反应(C-Skase)。分析了C-Skase与N-59单抗反应的表位结构域。结果表明，N-59mAb识别C-Skase分子中残基324-332(RDLYDPRDK)，最小表位区域为残基324-331(RDLYDPRD)。