Function of the tunnel in acetylcoenzyme A synthase/carbon monoxide dehydrogenase

Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry
Xiangshi TanPaul A Lindahl

Abstract

Acetylcoenzyme A synthase/carbon monoxide dehydrogenase (ACS/CODH) contains two Ni-Fe-S active-site clusters (called A and C) connected by a tunnel through which CO and CO2 migrate. Site-directed mutants A578C, L215F, and A219F were designed to block the tunnel at different points along the region between the two C-clusters. Two other mutant proteins F70W and N101Q were designed to block the region that connects the tunnel at the betabeta interface with a water channel also located at that interface. Purified mutant proteins were assayed for Ni/Fe content and examined by electron paramagnetic resonance spectroscopy. Analyses indicate that same metal clusters found in wild-type (WT) ACS/CODH (i.e., the A-, B-, C-, and probably D-clusters) are properly assembled in the mutant enzymes. Stopped-flow kinetics revealed that these centers in the mutants are rapidly reducible by dithionite but are only slowly reducible by CO, suggesting an impaired ability of CO to migrate through the tunnel to the C-cluster. Relative to the WT enzyme, mutant proteins exhibited little CODH or ACS activity (using CO2 as a substrate). Some ACS activity was observed when CO was a substrate, but not the cooperative CO inhibition effect characteristic of WT...Continue Reading

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Citations

May 22, 2009·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Brett M BarneyLance C Seefeldt
Sep 14, 2006·Journal of the American Chemical Society·Xiangshi TanPaul A Lindahl
Mar 2, 2010·Journal of the American Chemical Society·Liliya LundFrank M Raushel
Aug 8, 2008·Proceedings of the National Academy of Sciences of the United States of America·Fanny LerouxChristophe Léger
Mar 28, 2008·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Xiangshi Tan, Paul A Lindahl
Aug 16, 2017·Journal of Bacteriology·Amaya M Garcia CostasJohn W Peters
Feb 17, 2011·Faraday Discussions·Thomas LautierChristophe Léger
Oct 4, 2013·The Journal of Physical Chemistry. a·Aleksandra ChmielowskaMaria Jaworska
Apr 27, 2013·Chemical Reviews·Artur GoraJiri Damborsky
Sep 11, 2021·Archives of Biochemistry and Biophysics·David A KorasickJohn J Tanner
Jul 16, 2008·Chemical Reviews·Christophe Léger, Patrick Bertrand

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