Sagi I, Weinrich V, Levin I, Glotz C, Laschever M, Melamud M, Franceschi F, Weinstein S, Yonath A
Department of Structural Biology, Weizmann Institute, Rehovot, Israel.
Biophys Chem. 1995 Jun-Jul;55(1-2):31-41. doi: 10.1016/0301-4622(94)00140-f.
Crystals of various ribosomal particles, diffracting best to 2.9 A resolution were grown. Crystallographic data were collected from shock frozen crystals with intense synchrotron radiation at cryo temperature. For obtaining phase information, monofunctional reagents were prepared from an undecagold and a tetrairidium cluster, by attaching to them chemically reactive handles, specific for sulfhydryl moieties. Heavy-atom derivatives were prepared by a specific and quantitative binding of the undecagold cluster to an exposed sulfhydryl prior to the crystallization. To create potential binding sites on the halophilic and thermophilic ribosomal particles, which yield our best and most interesting crystals, exposed reactive moieties were inserted, using genetic and chemical procedures. In order to choose the appropriate locations for these insertions, the surfaces of the ribosomal particles were mapped by direct chemical determination of exposed amino and sulfhydryl groups.
生长出了各种核糖体颗粒的晶体，其衍射效果最佳可达2.9埃分辨率。在低温下，利用强同步辐射从快速冷冻的晶体中收集晶体学数据。为了获得相位信息，通过将对巯基部分具有化学反应活性的手柄化学连接到一个十一金簇和一个四铱簇上，制备了单功能试剂。在结晶之前，通过十一金簇与暴露的巯基的特异性定量结合制备重原子衍生物。为了在嗜盐和嗜热核糖体颗粒上创建潜在的结合位点（这些颗粒产生了我们最好且最有趣的晶体），使用遗传和化学方法插入了暴露的反应性基团。为了选择这些插入的合适位置，通过直接化学测定暴露的氨基和巯基来绘制核糖体颗粒的表面图谱。
