PMID- 8391727
OWN - NLM
STAT- MEDLINE
DCOM- 19930805
LR  - 20091119
IS  - 0340-6245 (Print)
IS  - 0340-6245 (Linking)
VI  - 69
IP  - 5
DP  - 1993 May 3
TI  - Properties of chimeric (tissue-type/urokinase-type) plasminogen activators 
      obtained by fusion at the plasmin cleavage site.
PG  - 466-72
AB  - Two hybrid plasminogen activators (K2tu-PA and FK2tu-PA), linking the kringle 2 
      domain or the finger plus the kringle 2 domains of tissue-type plasminogen 
      activator (t-PA) to the catalytic domain of single-chain urokinase-type 
      plasminogen activator (scu-PA) were studied. At variance with similar constructs 
      previously reported, they were obtained by fusion of the t-PA and scu-PA derived 
      portions at their plasmin cleavage site (between Arg275 of t-PA and Ile159 of 
      scu-PA), thus eliminating from scu-PA the two peptide bonds (Glu143-Leu144 and 
      Arg156-Phe157) that lead to low molecular weight scu-PA and to 
      thrombin-inactivated tcu-PA. The specific activities of K2tu-PA and FK2tu-PA, as 
      measured by fibrin plate were 2.5 x 10(6) and 1.0 x 10(6) t-PA equivalent 
      units/mg, respectively. Activation of plasminogen by hybrid PAs was stimulated by 
      both CNBr-digested fibrinogen (40- and 80-fold) and Des-A-fibrin monomers (6- and 
      12-fold). The relatively weak stimulation of chimeric PAs by minimally degraded 
      fibrin monomers was consistent with their reduced fibrin binding capacity. Like 
      scu-PA, the chimeric PAs, in the single-chain form, were insensitive to 
      inhibition, as they retained full activity after prolonged incubation in plasma 
      and did not interact with SDS-reactivated recombinant PAI-1. The concentration 
      producing 50% lysis of blood clots in 3 h was 0.5 microgram/ml for K2tu-PA and 1 
      microgram/ml for FK2tu-PA, as compared to 0.5 microgram/ml and > 2 micrograms/ml 
      for t-PA and scu-PA, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
FAU - Colucci, M
AU  - Colucci M
AD  - Dipartimento di Scienze Biomediche e Oncologia Umana, Universita di Bari, Italy.
FAU - Cavallo, L G
AU  - Cavallo LG
FAU - Agnelli, G
AU  - Agnelli G
FAU - Mele, A
AU  - Mele A
FAU - Burgi, R
AU  - Burgi R
FAU - Heim, J
AU  - Heim J
FAU - Semeraro, N
AU  - Semeraro N
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Thromb Haemost
JT  - Thrombosis and haemostasis
JID - 7608063
RN  - 0 (Fibrinolytic Agents)
RN  - 0 (Peptide Fragments)
RN  - 0 (Plasminogen Activator Inhibitor 1)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 9001-31-4 (Fibrin)
RN  - EC 3.4.21.- (Plasminogen Activators)
RN  - EC 3.4.21.68 (Tissue Plasminogen Activator)
RN  - EC 3.4.21.7 (Fibrinolysin)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Drug Design
MH  - Fibrin/pharmacology
MH  - Fibrinolysin/metabolism
MH  - Fibrinolysis/drug effects
MH  - Fibrinolytic Agents
MH  - Humans
MH  - Peptide Fragments/genetics
MH  - Plasminogen Activator Inhibitor 1/metabolism
MH  - Plasminogen Activators/metabolism
MH  - Protein Conformation
MH  - *Protein Engineering
MH  - Recombinant Fusion Proteins/genetics/*pharmacology
MH  - Tissue Plasminogen Activator/genetics/*pharmacology
MH  - Urokinase-Type Plasminogen Activator/genetics/*pharmacology
EDAT- 1993/05/03 00:00
MHDA- 1993/05/03 00:01
CRDT- 1993/05/03 00:00
PHST- 1993/05/03 00:00 [pubmed]
PHST- 1993/05/03 00:01 [medline]
PHST- 1993/05/03 00:00 [entrez]
PST - ppublish
SO  - Thromb Haemost. 1993 May 3;69(5):466-72.