[Molecular cloning and expression of the cDNA encoding angiogenesis inhibitor K4K5 with Pichia pastoris]

Sheng Wu Gong Cheng Xue Bao. 2001 Mar;17(2):126-30.
[Article in Chinese]

Abstract

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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Angiogenesis Inhibitors / genetics*
  • Animals
  • Cattle
  • Cell Division / drug effects
  • Chick Embryo
  • Cloning, Molecular
  • DNA, Complementary / isolation & purification*
  • Endothelium, Vascular / drug effects
  • Kringles*
  • Pichia / genetics*
  • Plasminogen / genetics*
  • Recombinant Proteins / biosynthesis

Substances

  • Angiogenesis Inhibitors
  • DNA, Complementary
  • Recombinant Proteins
  • Plasminogen