Kruft V, Wittmann-Liebold B
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, Federal Republic of Germany.
Biochemistry. 1991 Dec 24;30(51):11781-7. doi: 10.1021/bi00115a007.
Limited proteolysis was used in combination with two-dimensional gel electrophoresis, blotting, and amino acid sequence analysis to investigate the surface of intact ribosomal subunits at the peptide and amino acid level. Surface sites of 14 ribosomal proteins from Escherichia coli 50S subunits were determined using proteases with different specificities. To assess the evolutionary conservation of ribosomal topography among eubacteria, large subunits from Bacillus stearothermophilus were also subjected to limited proteolysis. The results obtained indicate a conservation of the three-dimensional ribosomal structure at the peptide level. The data for the eubacterial ribosomes are in full agreement with the model of the 50S protein topography derived from immunological data. Furthermore, peptide surface regions of archaebacterial ribosomes have been investigated. The results presented in this work prove that limited proteolysis can successfully be applied to halophilic and thermophilic ribosomes from archaebacteria.
有限蛋白酶解与二维凝胶电泳、印迹法及氨基酸序列分析相结合，用于在肽和氨基酸水平上研究完整核糖体亚基的表面。使用具有不同特异性的蛋白酶确定了来自大肠杆菌50S亚基的14种核糖体蛋白的表面位点。为评估真细菌中核糖体拓扑结构的进化保守性，嗜热脂肪芽孢杆菌的大亚基也进行了有限蛋白酶解。所得结果表明在肽水平上核糖体三维结构具有保守性。真细菌核糖体的数据与源自免疫学数据的50S蛋白拓扑结构模型完全一致。此外，还研究了古细菌核糖体的肽表面区域。这项工作中呈现的结果证明有限蛋白酶解可成功应用于来自古细菌的嗜盐和嗜热核糖体。
