کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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2021176 | 1069234 | 2008 | 7 صفحه PDF | دانلود رایگان |
Mammalian target of rapamycin (TOR) controls cell growth and metabolism in response to the availability of nutrients, growth factors, and the cellular energy status. Misregulation of TOR can result in a pathogenic increase or decrease in organ size and in cancer. TOR can be inhibited by binding of a complex of rapamycin and FKBP to the FKBP-rapamycin binding (FRB) domain. Rapamycin and derivatives of it have been used as immunosuppressive drugs. Because TOR is further an interesting drug target in cancer research, we established an expression, purification, and refolding protocol for the FRB domain of human TOR (hFRB). hFRB is extracted from inclusion bodies, purified by reversed phase HPLC, and refolded by drop-wise dilution of the denatured protein into a native buffer. The procedure is very simple and can easily be scaled up to prepare large amounts of functional protein for high-throughput cancer drug screening assays by NMR and other techniques.
Journal: Protein Expression and Purification - Volume 59, Issue 1, May 2008, Pages 31–37