Open AccessCloning, purification and preliminary crystallographic analysis of cobalamin methyltransferases from Rhodobacter capsulatus
Author(s) -
Seyedarabi Arefeh,
Hutchison Thomas,
To Teng Teng,
Deery Evelyne,
Brindley Amanda,
Warren Martin J.,
Pickersgill Richard W.
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110042910
Subject(s) - rhodobacter , cobalamin , methyltransferase , decarboxylation , chemistry , stereochemistry , methylation , corrin , crystallography , vitamin b12 , biochemistry , catalysis , gene , mutant
Of the 30 biosynthetic steps necessary for the production of cobalamin (vitamin B 12 ), eight involve the addition of S ‐adenosylmethionine‐derived methyl groups to the tetrapyrrole framework. These eight methyl additions are catalysed by six canonical methyltransferase domains and one noncanonical methyltransferase domain. Recombinant forms of four methyltransferases from Rhodobacter capsulatus , CobJ, CobM, CobF and CobL, and of the C‐terminal noncanonical domain of CobL (CobL‐C) have been crystallized, some in more than one crystal form. Most of the crystals diffracted to beyond 2.5 Å resolution and all are suitable for structure determination. Crystals of CobM and CobJ, which are involved in ring contraction, and of CobL, which is involved in two methylations and decarboxylation, are reported for the first time.