PMID- 8449917
OWN - NLM
STAT- MEDLINE
DCOM- 19930415
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 268
IP  - 8
DP  - 1993 Mar 15
TI  - The tissue plasminogen activator finger domain confers fibrin-dependent 
      enhancement of catalytic activity to single-chain urokinase-type plasminogen 
      activator.
PG  - 5550-6
AB  - To determine whether the fibrin-binding domains of tissue plasminogen activator 
      (tPA) can confer enhanced catalytic activity to single-chain urokinase-type 
      plasminogen activator (scuPA), we constructed, expressed, and characterized the 
      kinetics of five recombinant tPA/scuPA hybrid molecules. The hybrid molecules 
      are: 1) tPA3-50 (tPA finger)/scuPA138-411, 2) tPA177-256 (tPA 
      kringle2)/scuPA140-411 (scuPA catalytic), 3) tPA3-50/tPA177-256/scuPA140-411, 4) 
      scuPA1-47 (scuPA growth factor)/tPA177-256/scuPA140-411, and 5) scuPA1-138 (scuPA 
      growth factor and kringle)tPA127-256/scuPA139-411. The amidolytic activity of all 
      hybrids was comparable, as were the kinetics for conversion from single-chain to 
      two-chain plasminogen activator. We found that 1) the lag time prior to achieving 
      maximal velocity among these hybrids varied, 2) hybrids 2, 3, 4, and 5 were 
      2-134-fold more potent (by kcat/Km) than hybrid 1, and 3) those hybrid proteins 
      containing the tPA finger domain (hybrids 1 and 3) gave a 2-fold increase in 
      catalytic efficiency in the presence of DESAFIB (reptilase-digested fibrinogen). 
      These kinetic differences are likely mediated by changes in the tertiary 
      structure of the scuPA catalytic domain resulting from interactions between 
      catalytic and noncatalytic domains in the presence of fibrin.
FAU - Lubin, I M
AU  - Lubin IM
AD  - Division of Cardiology, Emory University School of Medicine, Atlanta, Georgia 
      30322.
FAU - Caban, R
AU  - Caban R
FAU - Runge, M S
AU  - Runge MS
LA  - eng
GR  - HL02414/HL/NHLBI NIH HHS/United States
GR  - HL08439-01/HL/NHLBI NIH HHS/United States
GR  - HL44307/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Recombinant Fusion Proteins)
RN  - 9001-31-4 (Fibrin)
RN  - 9001-32-5 (Fibrinogen)
RN  - 9001-91-6 (Plasminogen)
RN  - 9007-49-2 (DNA)
RN  - EC 3.4.21.68 (Tissue Plasminogen Activator)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Catalysis
MH  - Cell Line
MH  - DNA
MH  - Fibrin/metabolism
MH  - Fibrinogen/metabolism
MH  - Kinetics
MH  - Molecular Sequence Data
MH  - Plasminogen/metabolism
MH  - Recombinant Fusion Proteins/metabolism
MH  - Structure-Activity Relationship
MH  - Tissue Plasminogen Activator/*metabolism
MH  - Urokinase-Type Plasminogen Activator/*metabolism
EDAT- 1993/03/15 00:00
MHDA- 1993/03/15 00:01
CRDT- 1993/03/15 00:00
PHST- 1993/03/15 00:00 [pubmed]
PHST- 1993/03/15 00:01 [medline]
PHST- 1993/03/15 00:00 [entrez]
AID - S0021-9258(18)53356-1 [pii]
PST - ppublish
SO  - J Biol Chem. 1993 Mar 15;268(8):5550-6.