Little J W, Hill S A
Proc Natl Acad Sci U S A. 1985 Apr;82(8):2301-5. doi: 10.1073/pnas.82.8.2301.
Many proteins are organized as a set of compact functional domains connected by flexible, exposed segments of the polypeptide chain. To study one of these connector regions, we isolated a series of functional in-frame deletions in the central portion of a specific DNA-binding protein, the LexA repressor of Escherichia coli. These mutant proteins fell into two main classes: those with small deletions of two to eight amino acids functioned as repressor about as well as did wild type, while those with large deletions of 17-22 amino acids functioned well only at considerably higher concentrations. The mutant proteins were resistant to the specific cleavage reaction that triggers the SOS response. These data suggest that the conformation of the hinge region in LexA protein is important for cleavage. By contrast, the hinge plays a topological role in repressor function, connecting the two functional halves of the protein; in the SOS response, this function of the hinge is inactivated by cleavage, leading to inactivation of the repressor.
许多蛋白质由一组紧密的功能结构域组成,这些结构域由多肽链中灵活、暴露的片段连接。为了研究其中一个连接区域,我们在一种特定的DNA结合蛋白——大肠杆菌的LexA阻遏物的中央部分分离出了一系列功能性的读框内缺失。这些突变蛋白主要分为两类:那些缺失两到八个氨基酸的小缺失突变蛋白,其作为阻遏物的功能与野生型相当,而那些缺失17 - 22个氨基酸的大缺失突变蛋白,只有在相当高的浓度下才能正常发挥作用。这些突变蛋白对触发SOS反应的特异性切割反应具有抗性。这些数据表明,LexA蛋白中铰链区的构象对切割很重要。相比之下,铰链在阻遏物功能中起拓扑作用,连接蛋白质的两个功能半区;在SOS反应中,铰链的这一功能通过切割而失活,导致阻遏物失活。