期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1972
卷号：69
期号：12
页码：3638-3642
DOI：10.1073/pnas.69.12.3638
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：The allosteric enzyme aspartate transcarbamoylase (EC 2.1.3.2 ) was previously shown to consist of two functionally distinct types of polypeptide chains. X-ray diffraction and chemical studies showed that there are six copies of both catalytic (C) and regulatory (R) chains, and that the intact molecular complex (C6R6) has D3 symmetry. Organomercurials react preferentially with the four thiol groups on each R chain, dissociating the molecular complex. We show that 2-chloromercuri-4-nitrophenol reacts specifically and rapidly with the single C-chain thiol, which is believed to be near the catalytic site. This reaction inactivates the enzyme in solution and does not dissociate the molecular complex. Spectrophotometric titration and mercury analysis indicates that six molecules of this mercurial are firmly bound to the enzyme (R6C6), and crystallographic studies establish that only six sites, related by D3 symmetry, are modified. The known low reactivity of this C-chain thiol with other sulfhydryl reagents, the unusual structural requirements in the reaction with 2-chloromercuri-4-nitrophenol, and the spectral properties of the resulting derivative provide insight into the environment of this thiol. Probably, at least one positively charged group of the enzyme is nearby, and the environment of this thiol is at least partially hydrophobic.
关键词：enzyme activity ; sulfhydryl group ; allosteric enzyme ; mercurials
