Some inhibitors have bad solubility but low Ki (less then 1 uM), we keep inhibitors solid in the soaking drops and don't know the exact concentration. For inhibitors with good solubility, we keep the ration at 10 or higher, unfortunately, Ki of these inhibitors are not good, for example, around 30 uM.
----- 原始邮件 ----- 发件人: "Jürgen Bosch" <jubo...@jhsph.edu> 收件人: "胡小鹏" <huxp...@mail.sysu.edu.cn> 抄送: CCP4BB@JISCMAIL.AC.UK 发送时间: 星期六, 2010年 8 月 28日 下午 8:00:46 主题: Re: [ccp4bb] Problem with getting protein-inhibitor complex structure What concentration/ratio of your inhibitor did you use ? Also check this out: (short soaks, see time course experiment) Bosch et al. Using fragment cocktail crystallography to assist inhibitor design of Trypanosoma brucei nucleoside 2-deoxyribosyltransferase. J Med Chem (2006) vol. 49 (20) pp. 5939-46 Jürgen - Jürgen Bosch Johns Hopkins Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Phone: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-3655 http://web.mac.com/bosch_lab/ On Aug 28, 2010, at 1:40 AM, 胡小鹏 wrote: Dear all, We are working on protein-inhibitor complex crystal structures, although we did get nice crystals and soaked them with inhibitor for several days, all structures we obtained were empty!!There are nothing at the active site. For several crystals. their color turned very clearly from colorless to yellow the color of the inhibitors, nothing found either. These inhibitors work very well in enzyme assay. We, especially our students, are very depressed... Any suggestions will be really appreciated! Xiaopeng Hu, D.Sc., School of Pharmaceutical Sciences Sun Yat-sen University, Higher Education Mega Center Guangzhou, Guangdong, China 510006 Tel. +86 020 39943032 -- -- 胡小鹏 理学博士 教授 中山大学药学院结构生物学实验室 广东省 广州市 番禺区 大学城 510006 Tel: 020-39943032 Xiaopeng Hu, D.Sc., Professor School of Pharmaceutical Sciences Sun Yat-sen University, Higher Education Mega Center Guangzhou, Guangdong, China 510006 Tel. +86 020 39943032
