The Protein Journal

, Volume 29, Issue 3, pp 167–173

Expression and Purification of Functional Epitope of Pigment Epithelium-Derived Factor in E. coli with Inhibiting Effect on Endothelial Cells

  • Qing Gong
  • Xia Yang
  • Weibin Cai
  • Guoquan Gao
  • Zhonghan Yang
Article

DOI: 10.1007/s10930-010-9236-6

Cite this article as:
Gong, Q., Yang, X., Cai, W. et al. Protein J (2010) 29: 167. doi:10.1007/s10930-010-9236-6

Abstract

PEDF34, a functional epitope of pigment epithelium-derived factor (PEDF), obtained by chemical synthesis previously, shows potential anti-angiogenesis activity described before. We perform a novel method in this study for the expression and purification of recombinant PEDF34 in E. coli, and make it convenient, soluble and high yield to obtain this small peptide of PEDF. Human PEDF34 gene was cloned into the fusion-protein expression vector pGEX-4T-1, and the recombinant plasmid was transformed into E. coli strain BL21-DE3. GST-PEDF34 fusion protein was expressed, purified using chromatograph and identified by Western blotting. The purified fusion protein was digested by thrombin, and the small PEDF34 peptide was isolated by ultrafiltration. Circular dichroism (CD) analysis identified that secondary structure of PEDF34 mainly characterizes as α-helix. The 34-AA small peptide could cell-type-specifically inhibit viability of HUVECs in a dose-dependent manner and induce apoptosis of HUVECs. These results suggested that this type of recombinant PEDF34 may have potential in the treatment of angiogenesis-related diseases such as solid tumor.

Keywords

PEDF Epitope Angiogenesis Apoptosis HUVECs 

Abbreviations

PEDF

Pigment epithelium-derived factor

HUVECs

Human umbilical vein endothelial cells

LR

Laminin receptor

PNPLA2

Phospholipase A2/nutrin/patatin-like phospholipase domain-containing 2

GST

Glutathione-S-transferase

PI

Propidium iodide

E. coli

Escherchia coli

CD

Circular dichroism

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Qing Gong
    • 1
  • Xia Yang
    • 1
  • Weibin Cai
    • 1
  • Guoquan Gao
    • 1
  • Zhonghan Yang
    • 1
  1. 1.Department of Biochemistry, Zhongshan School of MedicineSun Yat-sen UniversityGuangzhouChina

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