[Molecular cloning and expression of the cDNA encoding angiogenesis inhibitor K4K5 with Pichia pastoris].
Kringles of human plasminogen except kringle 4 can inhibit the endothelial cell growth. To determine whether recombinant plasminogen kringle 4-5 (r-K4K5) can inhibit the growth of bovine capillary endothelial (BCE) cell and the angiogenesis of chick embryo chorioallantoic membrane (CAM), we cloned and constructed the modified cDNA region encoding kringles 4 and 5 (K4K5) of human plasmin (ogen) and expressed it with pichia multi-copy expression system. One clone, with the most productive expression was selected from hundreds of transformants. Our data showed that the expression product r-K4K5 (MW 21.5 kD) amounted to 150-250 mg/liter, over 80% of the total secreted protein. It could, dose-dependently, inhibit BCE cell proliferation and inhibit chick embryo CAM angiogenesis.
Guan
X Q
XQ
Department of Molecular Genetics, School of Basic Medical Sciences, Shanghai Medical University, Shanghai 200032, China.
Wang
Y X
YX
Wu
L C
LC
Song
H Y
HY
D020533:Angiogenesis Inhibitors / Q000235:genetics*; D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002642:Chick Embryo; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification*; D004730:Endothelium, Vascular / Q000187:drug effects; D018082:Kringles; D010843:Pichia / Q000235:genetics*; D010958:Plasminogen / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis
D020533:Angiogenesis Inhibitors; D018076:DNA, Complementary; D011994:Recombinant Proteins; D010958:Plasminogen