Acid native polyacrylamide gel electrophoresis:a method for studying the mechanism of action of HIV entry inhibitor ADS-J1
- VernacularTitle:酸性天然凝胶电泳法研究HIV进入抑制剂ADS-J1的作用机制
- Author:
Qinchao MAO
;
Hongtao WANG
;
Xugui LI
;
Chenglai XIA
;
Shibo JIANG
;
Shuwen LIU
- Publication Type:Journal Article
- Keywords:
HIV entry inhibitor;
gp41;
acid native polyacrylamide gel electrophoresis (AN-PAGE);
ADS-J1;
Type I enveloped viruses;
six-helix bundle
- From:
Chinese Pharmacological Bulletin
2010;26(1):25-28
- CountryChina
- Language:Chinese
-
Abstract:
Aim ADS-J1 is a low molecular HIV entry inhibitor targeting HIV transmembrane subunit gp41 through virtual screening from a compound library containing 20 000 molecules.This study is to investigate the binding sites of ADS-J1 on gp41.Methods Acid native polyacrylamide gel electrophoresis (AN-PAGE) assay was applied to test the binding ability of ADS-J1 with the peptides derived from gp41 N-terminal heptad repeat (NHR) region.Results It was reported previously that ADS-J1 could block the gp41 six-helix bundle (6-HB) formation using native polyacrylamide gel electrophoresis (N-PAGE).However,the binding sites could not be found because positive charged N-peptides derived from gp41 NHR could not show bands on the gel.In the present study,the AN-PAGE assay which could show N-peptides in the gel was established,and it was found that ADS-J1 could inhibit the gp41 6-HB formation.Moreover,ADS-J1 bound directly to the gp41 cavity region of NHR.The positively charged residue (K574) located in this region was critical for the binding of ADS-J1.Conclusions ADS-J1 inhibits HIV entry by targeting the cavity region of gp41 NHR,whereas K574 in the cavity plays a critical role for the binding.Furthermore,the AN-PAGE assay provides a simple method for studying the mechanism of action of virus entry inhibitors targeting the transmembrane protein of type I enveloped virus.
