Crystal structures of cytochrome c(L) and methanol dehydrogenase from Hyphomicrobium denitrificans: structural and mechanistic insights into interactions between the two proteins

Biochemistry
Masaki NojiriShinnichiro Suzuki

Abstract

Methanol dehydrogenase (Hd-MDH) and its physiological electron acceptor, cytochrome c(L) (Hd-Cyt c(L)), isolated from a methylotrophic denitrifying bacterium, Hyphomicrobium denitrificans A3151, have been kinetically and structurally characterized; the X-ray structures of Hd-MDH and Hd-Cyt c(L) have been determined using molecular replacement at 2.5 and 2.0 A resolution, respectively. To explain the mechanism for electron transfer between these proteins, the dependence of MDH activity on the concentration of Hd-Cyt c(L) has been investigated at pH 4.5-7.0. The Michaelis constant for Hd-Cyt c(L) shows the smallest value (approximately 0.3 microM) at pH 5.5. The pseudo-first-order rate constant (k(obs)) of the reduction of Hd-Cyt c(L) exhibits a hyperbolic concentration dependence of Hd-MDH at pH 5.5, although k(obs) linearly increases at pH 6.5. These findings indicate formation of a transient complex between these proteins during an electron transfer event. Hd-MDH (148 kDa) is a large tetrameric protein with an alpha(2)beta(2) subunit composition, showing a high degree of structural similarity with other MDHs. Hd-Cyt c(L) (19 kDa) exhibiting the alpha-band at 550.7 nm has a unique C-terminal region involving a disulfide bond be...Continue Reading

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Citations

Apr 21, 2011·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Jin Myung ChoiSung Haeng Lee
Aug 7, 2012·Biochemistry. Biokhimii︠a︡·A R GvozdevR I Gvozdev
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Mar 2, 2011·Biochemical and Biophysical Research Communications·Jie LiZong-Xiang Xia
Dec 21, 2014·Applied and Environmental Microbiology·Ming L WuLaura van Niftrik
Sep 27, 2018·IUBMB Life·Giovanna De SimonePaolo Ascenzi
Mar 29, 2018·MBio·Yue ZhengLudmila Chistoserdova
Aug 23, 2018·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Yue Wen DengAmy C Rosenzweig
Oct 20, 2020·ChemCatChem·Jorge NevarezRobert P Hausinger
Jul 7, 2020·Journal of Microbiology and Biotechnology·Suparna GhoshSung Haeng Lee
Feb 20, 2021·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Pedro D Sarmiento-Pavía, Martha E Sosa-Torres

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