Daas P J, Gerrits K A, Keltjens J T, van der Drift C, Vogels G D
Department of Microbiology, Faculty of Science, University of Nijmegen, The Netherlands.
J Bacteriol. 1993 Mar;175(5):1278-83. doi: 10.1128/jb.175.5.1278-1283.1993.
Methanol:5-hydroxybenzimidazolylcobamide methyltransferase (MT1) is the first of two enzymes required for transfer of the methyl group of methanol to 2-mercaptoethanesulfonic acid in Methanosarcina barkeri. MT1 binds the methyl group of methanol to its corrinoid prosthetic group only when the central cobalt atom of the corrinoid is present in the highly reduced Co(I) state. However, upon manipulation of MT1 and even during catalysis, the enzyme becomes inactivated as the result of Co(I) oxidation. Reactivation requires H2, hydrogenase, and ATP. Ferredoxin stimulated the apparent reaction rate of methyl group transfer. Here we report that one more protein fraction was found essential for the overall reaction and, more specifically, for formation of the methylated MT1 intermediate. The more of the protein that was present, the shorter the delay of the start of methyl group transfer. The maximum velocity of methyl transfer was not substantially affected by these varying amounts of protein. This demonstrated that the protein was involved in the activation of MT1. Therefore, it was called methyltransferase activation protein.
5-羟基苯并咪唑基钴胺素甲基转移酶(MT1)是巴氏甲烷八叠球菌中,将甲醇甲基转移至2-巯基乙烷磺酸所需的两种酶中的第一种。只有当类咕啉的中心钴原子处于高度还原的Co(I)状态时,MT1才会将甲醇的甲基结合到其类咕啉辅基上。然而,在对MT1进行操作时,甚至在催化过程中,由于Co(I)氧化,该酶会失活。重新激活需要H2、氢化酶和ATP。铁氧化还原蛋白刺激了甲基转移的表观反应速率。在此我们报告,发现另一种蛋白质组分对于整个反应,更具体地说,对于甲基化MT1中间体的形成是必不可少的。该蛋白质存在的量越多,甲基转移开始的延迟就越短。这些不同量的蛋白质对甲基转移的最大速度没有实质性影响。这表明该蛋白质参与了MT1的激活。因此,它被称为甲基转移酶激活蛋白。