Functional characterization of recombinant batroxobin, a snake venom thrombin-like enzyme, expressed from Pichia pastoris.
A thrombin-like enzyme of Bothrops atrox moojeni venom, batroxobin, specifically cleaves fibrinogen alpha chain, resulting in the formation of non-crosslinked fibrin clots. The cDNA encoding batroxobin was cloned, expressed in Pichia pastoris and the molecular function of purified recombinant protein was also characterized. The recombinant batroxobin had an apparent molecular weight of 33 kDa by SDS-PAGE analysis and biochemical activities similar to those of native batroxobin. The purified recombinant protein strongly converted fibrinogen into fibrin clot in vitro, and shortened bleeding time and whole blood coagulation time in vivo. However, it did not make any considerable alterations on other blood coagulation factors. Several lines of experimental evidence in this study suggest that the recombinant batroxobin is a potent pro-coagulant agent.
You
Weon-Kyoo
WK
R&D Center, Biobud Co. Ltd., Seoul 120-110, Republic of Korea.
Choi
Won-Seok
WS
Koh
You-Seok
YS
Shin
Hang-Cheol
HC
Jang
Yangsoo
Y
Chung
Kwang-Hoe
KH
D000595:Amino Acid Sequence; D012103:Batroxobin / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D010843:Pichia / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013917:Thrombin / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
D018076:DNA, Complementary; D011994:Recombinant Proteins; D012103:Batroxobin; D014357:Trypsin; D013917:Thrombin