发明公开
- 专利标题： Active site labelling of plasminogen
- 专利标题（中）： 活性位点 - 马立氏纤溶酶原。
-
申请号： EP91112857.7申请日： 1991-07-31
-
公开(公告)号： EP0470494A1公开(公告)日： 1992-02-12
- 发明人: Shore, Joseph D., Dr. , Bock, Paul E. Dr. , Olson, Steven T., Dr. , Day, Duane E., Dr.
- 申请人： HENRY FORD HEALTH SYSTEM, d/b/a HENRY FORD HOSPITAL
- 申请人地址： 2799 West Grand Boulevard Detroit, Michigan 48202-2689 US
- 专利权人： HENRY FORD HEALTH SYSTEM, d/b/a HENRY FORD HOSPITAL
- 当前专利权人： HENRY FORD HEALTH SYSTEM, d/b/a HENRY FORD HOSPITAL
- 当前专利权人地址： 2799 West Grand Boulevard Detroit, Michigan 48202-2689 US
- 代理机构： Koepe, Gerd L., Dipl.-Chem.
- 优先权： US562702 19900806
- 主分类号： C12Q1/56
- IPC分类号： C12Q1/56 ; G01N33/86 ; C12N9/64
摘要：
A plasminogen, which has a label incorporated into an active site thereof such that said plasminogen is enzymatically and catalytically inactive upon its conversion to plasmin, and an enzymatically and catalytically inactive plasmin with a label incorporated into an active site thereof.
A method of making a labelled plasminogen which is enzymatically and catalytically inactive upon conversion to plasmin comprising:
a) forming a streptokinase plasminogen complex,
b) inhibiting the plasminogen by attaching a peptide chloromethyl ketone thereto,
c) adding a detectable molecule or marker to the peptide group derived from said peptide chloromethyl ketone thereby forming a labelled complex, and
d) dissociating the streptokinase from the labelled plasminogen thereby forming a labelled plasminogen which is enzymatically and catalytically inactive upon conversion to plasmin.
Preferably, the peptide chloromethyl ketone is [-(acetylthio)acetyl]-phenylalanine, phenylalanine, arginine, chloromethyl ketone and the detectable molecule or marker is a fluorophore acetamide attached to the thiol group thereof.
A method of making a labelled plasminogen which is enzymatically and catalytically inactive upon conversion to plasmin comprising:
a) forming a streptokinase plasminogen complex,
b) inhibiting the plasminogen by attaching a peptide chloromethyl ketone thereto,
c) adding a detectable molecule or marker to the peptide group derived from said peptide chloromethyl ketone thereby forming a labelled complex, and
d) dissociating the streptokinase from the labelled plasminogen thereby forming a labelled plasminogen which is enzymatically and catalytically inactive upon conversion to plasmin.
Preferably, the peptide chloromethyl ketone is [-(acetylthio)acetyl]-phenylalanine, phenylalanine, arginine, chloromethyl ketone and the detectable molecule or marker is a fluorophore acetamide attached to the thiol group thereof.
信息查询
