Detection of Bax protein conformational change using a surface plasmon resonance imaging-based antibody chip

被引:32
作者
Kim, M
Jung, SO
Park, K
Jeong, EJ
Joung, HA
Kim, TH
Seol, DW
Chung, BH
机构
[1] Korea Res Inst Biosci & Biotechnol, Bionanotechnol Res Ctr, Taejon 305600, South Korea
[2] Chosun Univ, Sch Med, Dept Biochem, Kwangju, South Korea
[3] Univ Pittsburgh, Sch Med, Dept Surg, Pittsburgh, PA 15261 USA
关键词
SPR imaging; conformational change; Bax; TRAIL; SPR sensor;
D O I
10.1016/j.bbrc.2005.10.155
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We describe an antibody chip technology that uses a surface plasmon resonance (SPR) imaging system to examine the conformational change of a protein. In this study, we used Bax protein, a pro-apoptotic member of the Bcl-2 family of proteins, as a model protein to investigate the conformational alteration triggered by a TNF-related apoptosis-inducing ligand (TRAIL), a potent inducer of apoptosis. To develop the antibody chip for detecting the Bax conformational change, we immobilized Bax. monoclonal antibody 6A7, which recognizes only a conformationally changed Bax protein on a gold surface. The resultant immobilized Bax antibodies provided specific and accurate measurements of the active conformation-specific epitope in the apoptotic cancer cells treated with the TRAIL; these measurements corresponded to the data obtained by immunoprecipitation analysis using an active conformation-specific Bax antibody (6A7). The results of our study indicated that TRAIL-induced Bax structural change could be monitored quickly and simply using an SPR imaging system, thus demonstrating the potential for using such a system for the analysis of conformational properties of target proteins. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:1834 / 1838
页数:5
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