PMID- 8117652
OWN - NLM
STAT- MEDLINE
DCOM- 19940404
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 33
IP  - 8
DP  - 1994 Mar 1
TI  - Design and evaluation of a thrombin-activable plasminogen activator.
PG  - p606-12
AB  - A new chimeric plasminogen activator with high fibrin affinity was designed to 
      bind fibrin and to initiate clot destruction, following activation by thrombin. 
      The chimeric activator, 59D8-scuPA-T, was made from the Fab fragment of an 
      anti-fibrin antibody (59D8) and a C-terminal portion of a thrombin-activable low 
      molecular weight single-chain urokinase plasminogen activator, scuPA-T, obtained 
      by deletion of Phe-157 and Lys-158 from low molecular weight single-chain 
      urokinase-type plasminogen activator (scuPA) by site-directed mutagenesis. The 
      chimeric molecule had a molecular mass of 91,000, a value consistent with one 
      59D8 light chain (M(r) = 27,000) and one 59D8 heavy-chain Fd fragment fused to 
      low molecular weight scuPA (M(r) = 64,000). According to its design, 59D8-scuPA-T 
      was activated by thrombin but not by plasmin, whereas the control chimeric 
      molecule, 59D8-scuPA, was activated by plasmin but not by thrombin. When 
      activated by thrombin, 59D8-scuPA-T converted plasminogen to plasmin. In vitro 
      plasma clot lysis assays showed that 59D8-scuPA-T lysed clots performed by 
      thrombin and that heparin and hirudin could prevent clot lysis. When incorporated 
      as part of a thrombin-induced clot, only 59D8-scuPA-T was able to lyse the clot 
      while 59D8-scuPA and high molecular weight scuPA were ineffective. Together these 
      results demonstrate that 59D8-scuPA-T is a thrombin-activable plasminogen 
      activator that offers selective thrombolysis of thrombin-rich clots over more 
      established, aged clots, and may also act as an antithrombotic agent.
FAU - Yang, W P
AU  - Yang WP
AD  - Department of Macromolecular Biochemistry, Bristol-Myers Squibb Pharmaceutical 
      Research Institute, Princeton, New Jersey 08543-4000.
FAU - Goldstein, J
AU  - Goldstein J
FAU - Procyk, R
AU  - Procyk R
FAU - Matsueda, G R
AU  - Matsueda GR
FAU - Shaw, S Y
AU  - Shaw SY
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Immunoglobulin Fab Fragments)
RN  - 0 (Oligodeoxyribonucleotides)
RN  - 0 (Recombinant Proteins)
RN  - 9001-91-6 (Plasminogen)
RN  - EC 3.4.21.5 (Thrombin)
RN  - EC 3.4.21.7 (Fibrinolysin)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Base Sequence
MH  - Blood Coagulation
MH  - Cell Line
MH  - Cloning, Molecular
MH  - Drug Design
MH  - Fibrinolysin/pharmacology
MH  - Fibrinolysis/drug effects
MH  - Humans
MH  - Immunoglobulin Fab Fragments
MH  - Molecular Sequence Data
MH  - Oligodeoxyribonucleotides
MH  - Plasminogen/metabolism
MH  - Recombinant Proteins/genetics/pharmacology
MH  - Thrombin/*pharmacology
MH  - Urokinase-Type Plasminogen Activator/*genetics/pharmacology
EDAT- 1994/03/01 00:00
MHDA- 1994/03/01 00:01
CRDT- 1994/03/01 00:00
PHST- 1994/03/01 00:00 [pubmed]
PHST- 1994/03/01 00:01 [medline]
PHST- 1994/03/01 00:00 [entrez]
AID - 10.1021/bi00174a043 [doi]
PST - ppublish
SO  - Biochemistry. 1994 Mar 1;33(8):p606-12. doi: 10.1021/bi00174a043.