Zabin I
Mol Cell Biochem. 1982 Nov 26;49(2):87-96. doi: 10.1007/BF00242487.
Studies on beta-galactosidase alpha-complementation are reviewed. The isolation and structure of two beta-galactosidase fragments that form an enzymically active complex are described. One of these is a cyanogen bromide peptide from whole beta-galactosidase; the other is a dimeric protein from a lacZ deletion mutant of Escherichia coli. The mechanism most likely involves an initial binding of two cyanogen bromide peptides to the dimer, followed by formation of a tetramer, and finally a slow conformational change of the complex to a native-like enzyme. The overall reaction is essentially irreversible. A region of the polypeptide chain involved in dimer-dimer contact must be supplied by the cyanogen bromide peptide. alpha-Complemented enzyme contains overlapping sequences. Proteolytic experiments were carried out to determine the origin of the functionally important segment. The effect on alpha-complementation of amino acid substitutions at four positions in the polypeptide chain was investigated. The implications of these results for beta-galactosidase structure and for proteins in general are discussed.
本文综述了β-半乳糖苷酶α-互补的研究。描述了形成具有酶活性复合物的两个β-半乳糖苷酶片段的分离和结构。其中一个是来自完整β-半乳糖苷酶的溴化氰肽;另一个是来自大肠杆菌lacZ缺失突变体的二聚体蛋白。其机制很可能涉及两个溴化氰肽首先与二聚体结合,随后形成四聚体,最后复合物缓慢构象变化为类似天然酶的形式。整个反应基本上是不可逆的。参与二聚体-二聚体接触的多肽链区域必须由溴化氰肽提供。α-互补酶包含重叠序列。进行了蛋白水解实验以确定功能重要片段的来源。研究了多肽链中四个位置的氨基酸取代对α-互补的影响。讨论了这些结果对β-半乳糖苷酶结构以及一般蛋白质的意义。