|Table of Contents|

Specific labeling of the cell membrane-bound estrogen receptor(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2014 02
Page:
227-232
Research Field:
Articles
Publishing date:

Info

Title:
Specific labeling of the cell membrane-bound estrogen receptor
Author(s):
YUAN Yun LIN Zhiwei GUO Jiajia WANG Fei YUAN Xiaohong
1College of Life Sciences, Southwest University of Science and Technology, Mianyang 621000, China 2Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China 3Chengdu University of Technology, Chengdu 610059, China
Keywords:
estrogen receptor phosphopantetheinyl transferase labeling coenzyme A
CLC:
Q291 : Q78
PACS:
DOI:
10.3724/SP.J.1145.2014.00227
DocumentCode:

Abstract:
Cell membrane-bound estrogen receptors (cmERs) mediate the non-genomic effects of estrogen, which is involved in the occurrence of breast cancer and many other estrogen-related diseases. However, the mechanism study is hindered by the lack of a specific labeling method which can efficiently differentiate between cmERs and cytoplasmic ERs. The purpose of this study was to develop the specific labeling of the cmER. We expressed and purified glutathione S-transferase fusion proteins which contained three different specific peptides A1, S6 and ybbR. Then we compared the labeling specificity and efficiency of the proteins by two different phosphopantetheinyl transferase (PPTase): AcpS and Sfp. A1 peptide, which was shown with higher labeling efficiency, was constructed into the C-terminus of the estrogen receptor α (ERα-A1) and then the ERα-A1 expression, its genomic and nongenomic function and labeling of biotin by AcpS were measured in cells. The results showed that coupling of A1 tag did not affect the ER protein expression or its activity. AcpS could specifically label biotin to cmERα-A1with the coenzyme A-biotin as a substrate. These results provide a new method for the study of cmER-mediated estrogen nongenomic signal transduction.

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Last Update: 2014-05-04