The AAA+ motor complex of subunits CobS and CobT of cobaltochelatase visualized by single particle electron microscopy

Lundqvist, Joakim, Elmlund, Dominika, Heldt, Dana, Deery, Evelyne, Söderberg, Christopher A.G., Hansson, Mats, Warren, Martin ORCID: https://orcid.org/0000-0002-6028-6456 and Al-Karadaghi, Salam (2009) The AAA+ motor complex of subunits CobS and CobT of cobaltochelatase visualized by single particle electron microscopy. Journal of Structural Biology, 167 (3). pp. 227-234. ISSN 1047-8477

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Abstract

Cobalamins belong to the tetrapyrrole family of prosthetic groups. The presence of a metal ion is a key feature of these compounds. In the oxygen-dependent (aerobic) cobalamin biosynthetic pathway, cobalt is inserted into a ring-contracted tetrapyrrole called hydrogenobyrinic acid a,c-diamide (HBAD) by a cobaltochelatase that is constituted by three subunits, CobN, CobS and CobT, with molecular masses of 137, 37 and 71 kDa, respectively. Based on the similarities with magnesium chelatase, cobaltochelatase has been suggested to belong to the AAA+ superfamily of proteins. In this paper we present the cloning of the Brucella melitensis cobN, cobS and cobT, the purification of the encoded protein products, and a single-particle reconstruction of the macromolecular assembly formed between CobS and CobT from negatively stained electron microscopy images of the complex. The results show for the first time that subunits CobS and CobT form a chaperone-like complex, characteristic for the AAA+ class of proteins. The molecules are arranged in a two-tiered ring structure with the six subunits in each ring organized as a trimer of dimers. The similarity between this structure and that of magnesium chelatase, as well as analysis of the amino acid sequences confirms the suggested evolutionary relationship between the two enzymes.

Item Type: Article
Additional Information: Funding Information: This work was supported by a grant from the Swedish Natural Science Research Council to S. Al-Karadaghi.
Uncontrolled Keywords: catalysis,cobalt,metallation,tetrapyrrole,vitamin b,structural biology ,/dk/atira/pure/subjectarea/asjc/1300/1315
Faculty \ School: Faculty of Science
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Depositing User: LivePure Connector
Date Deposited: 20 Sep 2022 15:31
Last Modified: 27 Sep 2022 08:37
URI: https://ueaeprints.uea.ac.uk/id/eprint/88529
DOI: 10.1016/j.jsb.2009.06.013

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