Bosch Jürgen, Robien Mark A, Mehlin Christopher, Boni Erica, Riechers Aaron, Buckner Frederick S, Van Voorhis Wesley C, Myler Peter J, Worthey Elizabeth A, DeTitta George, Luft Joseph R, Lauricella Angela, Gulde Stacey, Anderson Lori A, Kalyuzhniy Oleksandr, Neely Helen M, Ross Jenni, Earnest Thomas N, Soltis Michael, Schoenfeld Lori, Zucker Frank, Merritt Ethan A, Fan Erkang, Verlinde Christophe L M J, Hol Wim G J
Department of Biochemistry, Division of Infectious Disease, and Howard Hughes Medical Institute, University of Washington, Seattle, Washington 98195, USA.
J Med Chem. 2006 Oct 5;49(20):5939-46. doi: 10.1021/jm060429m.
The 1.8 A resolution de novo structure of nucleoside 2-deoxyribosyltransferase (EC 2.4.2.6) from Trypanosoma brucei (TbNDRT) has been determined by SADa phasing in an unliganded state and several ligand-bound states. This enzyme is important in the salvage pathway of nucleoside recycling. To identify novel lead compounds, we exploited "fragment cocktail soaks". Out of 304 compounds tried in 31 cocktails, four compounds could be identified crystallographically in the active site. In addition, we demonstrated that very short soaks of approximately 10 s are sufficient even for rather hydrophobic ligands to bind in the active site groove, which is promising for the application of similar soaking experiments to less robust crystals of other proteins.
通过单波长反常散射（SAD）相位法，已确定了布氏锥虫核苷2-脱氧核糖基转移酶（TbNDRT，EC 2.4.2.6）在无配体状态和几种配体结合状态下分辨率为1.8埃的从头结构。这种酶在核苷回收的补救途径中很重要。为了鉴定新型先导化合物，我们采用了“片段混合浸泡法”。在31种混合液中尝试的304种化合物中，有4种化合物可通过晶体学方法在活性位点鉴定出来。此外，我们证明，即使对于相当疏水的配体，大约10秒的极短浸泡时间也足以使其结合到活性位点凹槽中，这对于将类似的浸泡实验应用于其他蛋白质的稳定性较差的晶体很有前景。
