Endo S, Nagayama K, Wada A
Department of Physics, Faculty of Science, University of Tokyo, Japan.
J Biomol Struct Dyn. 1985 Oct;3(2):409-21. doi: 10.1080/07391102.1985.10508426.
Protease susceptibility of homologous proteins in their native conformations was studied. This work aims to establish a broad and quantitative basis for the utilization of protease digestion to analyze the local stability of native proteins. Using high-performance liquid chromatography (HPLC) the time course of the proteolytic degradation of intact proteins was quantitatively traced. Rapid separation of peptide fragments with HPLC made possible the elucidation of sequential digestion originating from the cleavage at a very few sites which are locally unstable in the protein structure. Using four serine proteases, chymotrypsin, trypsin, elastase and subtilisin BPN', we found some common trends in proteolysis for a group of proteins of the cytochrome c family. By comparing of the proteolysis and thermal denaturation with ten homologous cytochromes c extracted from horse, beef, Candida krusei, Saccharomyces cerevisiae, chicken, tuna, pigeon, rabbit, dog and rat, protease susceptibility was related to locally unfolding states intrinsic to the native conformation.
研究了同源蛋白质在其天然构象下对蛋白酶的敏感性。这项工作旨在为利用蛋白酶消化分析天然蛋白质的局部稳定性建立广泛而定量的基础。使用高效液相色谱（HPLC）对完整蛋白质的蛋白水解降解过程进行了定量追踪。HPLC对肽片段的快速分离使得阐明源自蛋白质结构中极少数局部不稳定位点的切割的顺序消化成为可能。使用四种丝氨酸蛋白酶，即胰凝乳蛋白酶、胰蛋白酶、弹性蛋白酶和枯草杆菌蛋白酶BPN'，我们发现细胞色素c家族一组蛋白质在蛋白水解方面存在一些共同趋势。通过比较从马、牛肉、克鲁斯假丝酵母、酿酒酵母、鸡、金枪鱼、鸽子、兔子、狗和大鼠中提取的十种同源细胞色素c的蛋白水解和热变性，蛋白酶敏感性与天然构象固有的局部解折叠状态相关。
