Development and Application of Fluorogenic Peptide Substrates for Determination fo Blood Clotting Proteases
凝血蛋白酶测定荧光肽底物的研制及应用
基本信息
- 批准号:61880016
- 负责人:
- 金额:$ 5.76万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Developmental Scientific Research
- 财政年份:1986
- 资助国家:日本
- 起止时间:1986 至 1987
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Seventy-four peptide amides of 7-amino-4-methylcounarine (Mec), which contain various Boc-x-y-Arg-NH-Mecs, were newly synthesized and tested to find specific substrates for blood clotting proteases and trypsin, The X and Y residues of these substrates have been replaced by 12 and 15 different samino acids, respectively. Among these peptides, the followings were found to be most sensitive substrates for individual enzymes: Boc-Asp(OBz1)-Pro- Arg-NH-Mec (Kcat = 160 s^<-1> , Km = 11 (micrn)M, Kcat/Km = 15,000,000 M^<-1> s^<-1> ) for human (alpha)-thrombin, Z- Glu-Gly-Arg-NH-Mec (Kcat = 19 s^<-1>, Km = 59 (micrn)M, Kcat/km = 320,000 M^<-1> s^<-1>) for bovine factor Xa, Boc-Ln-Gly-Arg-NH-Mec (Kcat = 5.8 s^<-1>, Km = 140 (micrn)m, Kcat/Km = 42,000) for bovine factor XIIa, Boc-Asp(OBzl)- Ala-Arg-NH-Mec (Kcat = 9.2 9.2 s^<-1> , Km = 120 (micrn)m, Kcat/Km = 77,000 M^<-1>s^<-1>) for bovine activated protein C, and Boc-Gly-Phe-Arg-NH-Mec (Kcat = 29 s^<-1>, Km = 230 (micrn)m, Kcat/Km = 130,000 M^<-1>s^<-1>) for bovine plasma kallikrein. Moreover, Boc-Glu(OBzl)-Ala-Arg-NH-Mec (kcat = 46 s^<-1>, km = 370 (micrn)m, kcat/Km = 120,000 M^<-1>s^<-1>) was newly found as a good substrate for human factor XIa, Bovine trypsin effectively hydrolyzed peptide-NH-Mecs containing Ala and Por at the P2 site. The most reactive substrate was Boc-Gln-Ala-Arg-NH-Mec (kcat = 120 s^<-1>, km = 6.0 (micrn)m, kcat/km = 20,000,000 M^<-1> s^<-1>),
新合成了74种含有多种Boc-x-y-Arg-NH-Mecs的7-氨基-4-甲基丙氨酸(Mec)肽酰胺,并对其进行了测试,以寻找凝血蛋白酶和胰蛋白酶的特异性底物,这些底物的X和Y残基分别被12种和15种不同的氨基酸取代。在这些肽中,发现以下是对单个酶最敏感的底物:Boc-Asp (OBz1) pro - Arg-NH-Mec (Kcat = 160年代^ < 1 >,公里= 11 (micrn) M, Kcat /公里= 15000000 ^ < 1 > s ^ < 1 >)为人类凝血酶(α),Z - Glu-Gly-Arg-NH-Mec (Kcat = 19 s ^ < 1 >,公里= 59 (micrn) M, Kcat /公里= 320000 ^ < 1 > s ^ < 1 >)为Xa牛因素,Boc-Ln-Gly-Arg-NH-Mec (Kcat = 5.8 s ^ < 1 >,千米= 140 (micrn) M, Kcat /公里= 42000)对牛因素XIIa Boc-Asp (OBzl)——Ala-Arg-NH-Mec (Kcat = 9.2 - 9.2 s ^ < 1 >,千米= 120 (micrn) M, Kcat /公里= 77000 ^ < 1 > s ^ < 1 >)牛活化蛋白C,boc - gly - pheg - arg - nh - mec (Kcat = 29 s^<-1>, Km = 230 (micrn)m, Kcat/Km = 130,000 m ^<-1>s^<-1>)。此外,新发现Boc-Glu(OBzl)-Ala-Arg-NH-Mec (kcat = 46 s^<-1>, km = 370 (micrn)m, kcat/ km = 120,000 m ^<-1>s^<-1>)是人因子XIa的良好底物,牛胰蛋白酶在P2位点有效水解含有Ala和Por的肽- nh - mes。反应性最强的底物为Boc-Gln-Ala-Arg-NH-Mec (kcat = 120 s^<-1>, km = 6.0 (micrn)m, kcat/km = 20,000,000 m ^<-1> s^<-1>);
项目成果
期刊论文数量(43)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Nakamura,T.: Eur.J.Biochem.154. 511-521 (1986)
Nakamura,T.:Eur.J.Biochem.154。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
川畑俊一郎: "血液凝固・線溶・補体系とプロテアーゼ 実験医学,5巻,908-915頁" 羊工社(東京), 7 (1987)
Shunichiro Kawabata:“血液凝固、纤维蛋白溶解、补体系统和蛋白酶。实验医学,第 5 卷,第 908-915 页”Yokosha(东京),7 (1987)
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Sueyoshi, T.: "Bovine High-molecular-weight kininogen; The Maino Acid Sequence. Positions of Carbohydrate Chains and Disulfide Bridges in the Heavy Chain Portion." J. Bio. Chem.262. 2768-2779 (1987)
Sueyoshi, T.:“牛高分子量激肽原;主酸序列。重链部分中碳水化合物链和二硫键的位置。”
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
IWANAGA Sadaaki其他文献
IWANAGA Sadaaki的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('IWANAGA Sadaaki', 18)}}的其他基金
Role of Limulus Hemocytes in the Biological Defense System.
鲎血细胞在生物防御系统中的作用。
- 批准号:
04404090 - 财政年份:1992
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for General Scientific Research (A)
Basic studies on Development of Anti-thrombotic Agents
抗血栓药物开发的基础研究
- 批准号:
04557015 - 财政年份:1992
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for Developmental Scientific Research (B)
Molecular Mechanism of Extrinsic Blood Coagulation Pathway
外源性凝血途径的分子机制
- 批准号:
03044113 - 财政年份:1991
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for international Scientific Research
Studies on the Activity Measurement for Blood Proteases using their Monoclonal Antibodies
用单克隆抗体测定血液蛋白酶活性的研究
- 批准号:
02557016 - 财政年份:1990
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for Developmental Scientific Research (B)
Mechanism of Hemolymph Coagulation System in Invertebrates
无脊椎动物血淋巴凝固系统的机制
- 批准号:
02454539 - 财政年份:1990
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Initiation Mechanism of Extrinsic Blood Coagulation System
外源性凝血系统的启动机制
- 批准号:
63044110 - 财政年份:1988
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for international Scientific Research
Development of Synthetic Fluorogenic Peptide Substrates for Blood Clotting Proteases
凝血蛋白酶合成荧光肽底物的开发
- 批准号:
63870017 - 财政年份:1988
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for Developmental Scientific Research
Hemolymph Coagulation and Defence Systems in Invertebrate Animals
无脊椎动物的血淋巴凝固和防御系统
- 批准号:
62480453 - 财政年份:1987
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Studies on Molecular Abnormality of Blood Coagulation and Fibrinolytic Factors
凝血及纤溶因子分子异常研究
- 批准号:
60480497 - 财政年份:1985
- 资助金额:
$ 5.76万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
相似海外基金
Lead identification and pre-clinical studies on allosteric inhibitors of coagulation factor XIa
凝血因子 XIa 变构抑制剂的先导化合物鉴定和临床前研究
- 批准号:
10722847 - 财政年份:2022
- 资助金额:
$ 5.76万 - 项目类别:
Lead identification and pre-clinical studies on allosteric inhibitors of coagulation factor XIa
凝血因子 XIa 变构抑制剂的先导化合物鉴定和临床前研究
- 批准号:
10369394 - 财政年份:2022
- 资助金额:
$ 5.76万 - 项目类别:
Development of Reversible Inhibitors of Factor XIa
XIa 因子可逆抑制剂的开发
- 批准号:
nhmrc : 1024982 - 财政年份:2012
- 资助金额:
$ 5.76万 - 项目类别:
Project Grants
CONFORMATIONAL STUDY OF BLOOD COAGULATION FACTOR XIA
凝血因子XIA的构象研究
- 批准号:
7598464 - 财政年份:2007
- 资助金额:
$ 5.76万 - 项目类别:
A NEW FACTOR XIA INHIBITOR RELEASED FROM PLATELETS
从血小板中释放出新的 XIA 因子抑制剂
- 批准号:
3050851 - 财政年份:1990
- 资助金额:
$ 5.76万 - 项目类别:
A NEW FACTOR XIA INHIBITOR RELEASED FROM PLATELETS
从血小板中释放出新的 XIA 因子抑制剂
- 批准号:
3050850 - 财政年份:1989
- 资助金额:
$ 5.76万 - 项目类别:
A NEW FACTOR XIA INHIBITOR RELEASED FROM PLATELETS
从血小板中释放出新的 XIA 因子抑制剂
- 批准号:
3050849 - 财政年份:1988
- 资助金额:
$ 5.76万 - 项目类别: