STRUCTURE OF RIBOSOMAL S 15 PROTEIN BINDING SITE RNA

来自B的S 15蛋白在16 S RNA中的最小结合位点。 嗜热脂肪菌已被鉴定，这是一个65个核苷酸的RNA 由三个螺旋组成，在三向连接处相遇。 的88 残基蛋白质在1 mM浓度下形成表现良好的复合物， 得到良好分辨光谱的最佳温度为45 ℃。 我们 有能力用13 C，15 N标记这个大的复合物， 2 H以各种不同的方式。 我们准备了四个样品 13 C标记的RNA，其中仅一种类型的核苷酸被13 C标记 在每一个样品中。 此外，还制备了15 N标记的RNA， 制备了15 N标记蛋白质的复合物。 我们正在 记录调查光谱，以获得初始分配， 复杂的挑战 我们准备了各种各样的 氘代RNA：3 '，4'，5 '，5”-d4-RNA和具有特定碱基的RNA 氘化 配合物的光谱大大简化， 我们相信，我们应该能够获得大部分的连续 复合物中RNA的碱基、H V和HT质子的分配 使用750 MHz的Homestructive光谱。

A minimal binding site in 16S RNA for the S 15 protein from B. stearothermophilus has been identified, which is a 65 nucleotide RNA composed of three helices that meet at a three-way junction. The 88 residue protein forms a well-behaved complex at I mM concentration, and the optimal temperature for well-resolved spectra is 45-C. We have the capability of labeling this large complex with 13C, 15N, and 2H in a variety of different ways. We have prepared four samples 13C-labeled RNA where only one type of nucleotide is labeled with 13C in each sample. In addition, 15N labeled RNA has been prepared, and a complex with 15N-labeled protein has been made. We are in the process of recording survey spectra to obtain initial assignments for this challenging complex. We have prepared a variety of specifically deuterated RNAs: 3',4',5',5" -d4-RNA, and RNA with specific base deuteration. The spectra of the complex are greatly simplified, and we believe that we should be able to obtain most of the sequential assignments of the base, H V and HT protons of the RNA in the complex using homonuclear spectra at 750 MHz.