Expression, purification and characterization of recombinant Jerdonitin, a P-II class snake venom metalloproteinase comprising metalloproteinase and disintegrin domains.
Jerdonitin is a P-II class snake venom metalloproteinase comprising metalloproteinase and disintegrin domains. In this study, we established a high-level expression system in Pichia pastoris and developed a purification strategy for the recombinant Jerdonitin. This recombinant Jerdonitin degraded fibrinogen at a level of activity comparable with its wild type. The effects of recombinant Jerdonitin on inhibiting ADP-induced human platelet aggregation were in a dose-dependent manner with an IC(50) of 248nM. In addition, we reported here that Jerdonitin can significantly inhibit the growth of several cell lines, including human liver cancer cells (Bel7402), human leukemia cells (K562) and human gastric carcinoma cells (BGC823). This study offers recombinant Jerdonitin that will be valuable for further functional and structural studies of Jerdonitin.
Zhu
Lili
L
State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, 155 Yang Qiao Xi Lu, Fuzhou, Fujian 350002, China. zhulili@fjirsm.ac.cn
Yuan
Cai
C
Chen
Zhuo
Z
Wang
Wanyu
W
Huang
Mingdong
M
D000818:Animals; D045744:Cell Line, Tumor; D004396:Coloring Agents; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D019483:Disintegrins / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006801:Humans; D045726:Metalloproteases / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology*; D010843:Pichia / Q000378:metabolism; D010957:Plasmids; D010974:Platelet Aggregation / Q000187:drug effects; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D012910:Snake Venoms / Q000737:chemistry / Q000201:enzymology*; D013778:Tetrazolium Salts; D013844:Thiazoles
D004396:Coloring Agents; D018076:DNA, Complementary; D019483:Disintegrins; D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D012910:Snake Venoms; D013778:Tetrazolium Salts; D013844:Thiazoles; D045726:Metalloproteases; C022616:thiazolyl blue