Purification and characterization of recombinant murine endostatin in E. coli.
- Author:
Weon Kyoo YOU
1
;
Seung Ho SO
;
Hyosil LEE
;
Sun Young PARK
;
Mi Ran YOON
;
Soo Ik CHANG
;
Hyun Kyung KIM
;
Young Ae JOE
;
Yong Kil HONG
;
Soo Il CHUNG
Author Information
1. Mogam Biotechnology Research Institute, Yongin-City, Kyonggi-Do, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
recombinant endostatin;
angiogenesis inhibition;
prokaryotic expression system
- MeSH:
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
- From:Experimental & Molecular Medicine
1999;31(4):197-202
- CountryRepublic of Korea
- Language:English
-
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.
