and Characterization of Kringle 1-4.5 Domains of Human Plasminogen
ZHOU Qing-Wei, XIE Jing-Li1,
XIN Li, XU Ren, YE Qing1, LI Zai-Ping, GAN Ren-Bao*
( Institute of Biochemistry and Cell Biology, Shanghai
Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai
200031, China;
1State Key Lab of Bioreactor of East China University of Science and
Te chnology, Shanghai 200237, China )
Abstract The cDNA encoding Kringle 1-4 and part of
Kringle 5 domains of human plasminogen (K1-4.5), obtained from HepG2 by RT-PCR,
was cloned into expression vector pHIL-S1. The recombinant plasmid pHIL-K1-4.5
was transformed into Pichi pastoris GS115 and the recombinant yeast was
induced to express the recombinant proteins by methanol. The expressed proteins
were purified by lysine affinity chromatography to a purity of 95%. The
recombinant K1-4.5 inhibited the growth of bovine capillary endothelial cells
(BAEC) stimulated by the basic fibroblast growth factor (bFGF), in a
dosage-dependent manner with a half maximal concentration of 2 mg/L. rhK1-4.5
also inhibited 40% of the BAEC migration stimulated by bFGF in the
concentration of 1 mg/L.
Key
words
Pichia pastoris; expression; purification; bioactivity
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