Exp Mol Med.  1999 Dec;31(4):197-202.

Purification and characterization of recombinant murine endostatin in E. coli

• You WK
• So SH
• Lee H
• Park SY
• Yoon MR
• Chang SI
• Kim HK
• Joe YA
• Hong YK
• Chung SI

Abstract

Endostatin, a carboxyl-terminal fragment of collagen XVIII is known as an anti-angiogenic agent, that specifically inhibits the proliferation of endothelial cell and the growth of several primary tumor. We report here the purification and characterization of the recombinant murine endostatin (rmEndostatin) which was expressed in a prokaryotic expression system. This rmEndostatin has similar physiochemical properties of yeast-produced recombinant endostatin, and it also specifically inhibits the proliferation and migration of bovine capillary endothelial cells stimulated by basic fibroblast growth factor. The biological activity of rmEndostatin was also shown by its anti-angiogenic ability on the chorioallantoic membrane of chick embryo in vivo. In this article, we demonstrate the refolding and purification of rmEndostatin, expressed using E. coli system, to a biologically active and soluble form. In addition, these results confirm the activity of endostatin as a potent anti-angiogenic agent. Copyright 2000 Academic Press.

Keyword

recombinant endostatin; angiogenesis inhibition; prokaryotic expression system

MeSH Terms

Angiogenesis Inhibitors/pharmacology*
Angiogenesis Inhibitors/isolation & purification
Angiogenesis Inhibitors/genetics*
Animal
Blotting, Western
Cattle
Cell Movement/drug effects
Chick Embryo
Chorion/pathology
Chorion/drug effects
Circular Dichroism
Collagen/pharmacology*
Collagen/isolation & purification
Collagen/genetics*
Electrophoresis, Polyacrylamide Gel
Endothelium, Vascular/drug effects
Endothelium, Vascular/cytology
Escherichia coli/genetics*
Fibroblast Growth Factor, Basic/pharmacology
Mice
Neovascularization, Physiologic/drug effects
Peptide Fragments/pharmacology*
Peptide Fragments/isolation & purification
Peptide Fragments/genetics*
Protein Folding
Recombinant Proteins/pharmacology
Recombinant Proteins/isolation & purification
Recombinant Proteins/genetics
Solubility
Yeasts/genetics