Molecular and enzymatic characterization of XMRV protease by a cell-free proteolytic analysis
详细信息 查看全文
- 作者：Satoko Matsunaga ; Tatsuya Sawasaki ; Hirotaka Ode ; Ryo Morishita ; Ayako Furukawa ; Ryuta Sakuma ; Wataru Sugiura ; Hironori Sato ; Masato Katahira ; Akifumi Takaori-Kondo ; Naoki Yamamoto ; Akihide Ryo
- 关键词：XMRV ; Protease ; Cell-free protein synthesis ; AlphaSceen
- 刊名：Journal of Proteomics
- 出版年：2012
- 出版时间：3 August, 2012
- 年：2012
- 卷：75
- 期：15
- 页码：4863-4873
- 全文大小：1744 K
文摘
Xenotropic murine leukemia virus-related virus (XMRV) is a virus generated under artificial conditions by the recombination of 2 murine leukemia virus (MLV) proviruses, PreXMRV-1 and PreXMRV-2, during the in vivo passage of human prostate cancer cells in athymic nude mice. The molecular etiology of XMRV infection has not been characterized and its implication in human prostate cancer progression remains equivocal. As a step toward resolving this issue we developed an in vitro enzymatic assay system to characterize XMRV protease (PR)-mediated cleavage of host-cell proteins. Enzymatically-active XMRV PR protein was synthesized using a wheat-germ cell-free system. By monitoring cleavage activity of XMRV PR by AlphaScreen and 2-color immunoblot analyses, we revealed that the catalytic activity of XMRV PR is selectively blocked by the HIV PR inhibitor, Amprenavir, and identified several human tumor suppressor proteins, including PTEN and BAX, to be substrates of XMRV PR. This system may provide an attractive means for analyzing the function of retrovirus proteases and provide a technology platform for drug screening.