PMID- 9005443
OWN - NLM
STAT- MEDLINE
DCOM- 19970129
LR  - 20190909
IS  - 0269-2139 (Print)
IS  - 0269-2139 (Linking)
VI  - 9
IP  - 2
DP  - 1996 Feb
TI  - Construction and structure-activity relationships of chimeric prourokinase 
      derivatives with intrinsic thrombin-inhibitory potential.
PG  - 213-23
AB  - The blood clotting enzyme thrombin plays a central role in the aetiology of 
      occlusive disorders such as stroke and acute myocardial infarction. During 
      fibrinolytic therapy with plasminogen activators, thrombin is neutralized by 
      anticoagulative drugs. In order to combine plasminogen-activating and 
      thrombin-inhibitory activities we constructed chimeric derivatives of recombinant 
      single-chain, urokinase-type plasminogen activator (rscu-PA) which comprise the 
      kringle and protease domain of rscu-PA fused via a linker sequence to a 
      thrombin-inhibitory domain. The inhibitory domain contains a sequence element 
      directed to the active site of thrombin and a sequence taken from either hirudin 
      or the human thrombin receptor both binding to the fibrinogen recognition site of 
      thrombin. Analysing different sets of point mutants showed that the linker 
      between the protease domain and the active site-directed sequence is contributing 
      significantly to the thrombin-inhibitory potential. Kinetic analysis of thrombin 
      inhibition revealed that most of the chimeras tested competitively inhibit the 
      thrombin-mediated cleavage of a peptide substrate in a concentration-dependent 
      manner; however, in two examples the insertion of one glycine residue into the 
      active site directed-sequence abolished the blockade of the active site. This 
      supports the conclusion that the chimeras with high thrombin-inhibitory potential 
      interact with the active site and the fibrinogen recognition site of thrombin.
FAU - Wnendt, S
AU  - Wnendt S
AD  - Department of Molecular Pharmacology, Aachen, Germany.
FAU - Janocha, E
AU  - Janocha E
FAU - Schneider, J
AU  - Schneider J
FAU - Steffens, G J
AU  - Steffens GJ
LA  - eng
PT  - Journal Article
PL  - England
TA  - Protein Eng
JT  - Protein engineering
JID - 8801484
RN  - 0 (Antithrombins)
RN  - 0 (Hirudins)
RN  - 0 (Receptors, Thrombin)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 9001-32-5 (Fibrinogen)
RN  - EC 3.4.21.5 (Thrombin)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Amino Acid Sequence
MH  - Antithrombins/chemistry/genetics/metabolism
MH  - Binding Sites
MH  - Blood Coagulation/drug effects
MH  - Cloning, Molecular
MH  - Escherichia coli/genetics
MH  - Fibrinogen/metabolism
MH  - Gene Expression/genetics
MH  - Hirudins/chemistry/genetics
MH  - Humans
MH  - Kinetics
MH  - Kringles/genetics
MH  - Molecular Sequence Data
MH  - Receptors, Thrombin/chemistry/genetics
MH  - Recombinant Fusion Proteins/*genetics/metabolism/pharmacology
MH  - Sequence Alignment
MH  - Thrombin/*antagonists & inhibitors/genetics/metabolism
MH  - Urokinase-Type Plasminogen Activator/chemistry/*genetics
EDAT- 1996/02/01 00:00
MHDA- 1996/02/01 00:01
CRDT- 1996/02/01 00:00
PHST- 1996/02/01 00:00 [pubmed]
PHST- 1996/02/01 00:01 [medline]
PHST- 1996/02/01 00:00 [entrez]
AID - 10.1093/protein/9.2.213 [doi]
PST - ppublish
SO  - Protein Eng. 1996 Feb;9(2):213-23. doi: 10.1093/protein/9.2.213.