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Cloning, purification and preliminary crystallographic analysis of cobalamin methyltransferases from Rhodobacter capsulatus - PubMed

  • ️Fri Jan 01 2010

Cloning, purification and preliminary crystallographic analysis of cobalamin methyltransferases from Rhodobacter capsulatus

Arefeh Seyedarabi et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010.

Abstract

Of the 30 biosynthetic steps necessary for the production of cobalamin (vitamin B12), 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.

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Figures

Figure 1
Figure 1

SDS–PAGE analysis of the purified R. capsulatus methyltransferases (a) CobJ, (b) CobF, (c) CobM, (d) CobL-C and (e) CobL as visualized by Coomassie Blue staining. The molecular masses of the marker proteins in the lanes marked M are given in kDa.

Figure 2
Figure 2

Crystals of R. capsulatus methyltransferases: (a) CobJ (type I), (b) CobF (type I), (c) CobM (type I) and (d) CobL. This is the first report of CobJ, CobM and CobL crystals; CobF crystals have been reported previously from a different species.

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