Abstract
Kinetic, spectroscopic, and structural analysis tested the hypothesis that a chain of residues connecting the 4'-aminopyrimidine N1' atoms of thiamin diphosphates (ThDPs) in the two active centers of the Escherichia coli pyruvate dehydrogenase complex E1 component provides a signal transduction pathway. Substitution of the three acidic residues (Glu(571), Glu(235), and Glu(237)) and Arg(606) resulted in impaired binding of the second ThDP, once the first active center was filled, suggesting a pathway for communication between the two ThDPs. 1) Steady-state kinetic and fluorescence quenching studies revealed that upon E571A, E235A, E237A, and R606A substitutions, ThDP binding in the second active center was affected. 2) Analysis of the kinetics of thiazolium C2 hydrogen/deuterium exchange of enzyme-bound ThDP suggests half-of-the-sites reactivity for the E1 component, with fast (activated site) and slow exchanging sites (dormant site). The E235A and E571A variants gave no evidence for the slow exchanging site, indicating that only one of two active sites is filled with ThDP. 3) Titration of the E235A and E237A variants with methyl acetylphosphonate monitored by circular dichroism suggested that only half of the active sites were...Continue Reading
References
Dec 27, 1996·The Journal of Biological Chemistry·J YiJ R Guest
Jan 3, 1997·Science·D KernG Hübner
Feb 10, 1998·Biochemistry·N NemeriaF Jordan
Oct 3, 1998·Acta Crystallographica. Section D, Biological Crystallography·A T BrüngerG L Warren
Oct 5, 2001·The Journal of Biological Chemistry·N NemeriaF Jordan
Apr 17, 2002·Biochemistry·Palaniappa ArjunanWilliam Furey
Dec 27, 2002·Biochemistry·Natalia NemeriaWilliam Furey
Mar 26, 2003·The Journal of Biological Chemistry·Ewa M CiszakMulchand S Patel
Oct 16, 2003·Journal of the American Chemical Society·Frank JordanWilliam Furey
Oct 30, 2004·Science·Frank Jordan
Oct 30, 2004·Science·René A W FrankRichard N Perham
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Sep 21, 2005·Accounts of Chemical Research·Frank JordanEduard Sergienko
Oct 18, 2006·Biochemistry·Franziska SeifertKai Tittmann
Dec 22, 2006·Proceedings of the National Academy of Sciences of the United States of America·Natalia NemeriaFrank Jordan
Mar 3, 2007·The Journal of Biological Chemistry·Jun LiDavid T Chuang
Apr 25, 2007·Nucleic Acids Research·Ian W DavisDavid C Richardson
Aug 25, 2007·Biochemistry·Natalia NemeriaFrank Jordan
May 30, 2009·The FEBS Journal·Natalia S NemeriaFrank Jordan
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor
Citations
Oct 16, 2013·Journal of Molecular Modeling·Nataraj S PagadalaDavid S Wishart
Sep 14, 2012·Biochemistry·Natalia NemeriaFrank Jordan
Feb 4, 2012·Journal of the American Chemical Society·Anand BalakrishnanFrank Jordan
Apr 29, 2014·The FEBS Journal·Leighanne A Brammer BastaCaren L Freel Meyers
Sep 6, 2013·The FEBS Journal·Victoria I BunikNikolay V Lukashev
Jun 14, 2016·Structure·Ehab N M JirgisEdward N Baker
Nov 1, 2013·Biochemistry·Xiang Sheng, Yongjun Liu
Apr 3, 2013·Biochemistry·Kathrin Schröder-TittmannKai Tittmann
Apr 13, 2013·The Journal of Biological Chemistry·Krishnamoorthy ChandrasekharWilliam Furey
May 7, 2014·The Journal of Biological Chemistry·Mulchand S PatelFrank Jordan
Sep 20, 2019·Nature·Shaobo DaiKai Tittmann
Sep 12, 2014·The Journal of Biological Chemistry·Palaniappa ArjunanWilliam Furey
Nov 7, 2019·Scientific Reports·Henry C Wilkinson, Paul A Dalby
Mar 7, 2020·Scientific Reports·Henry C Wilkinson, Paul A Dalby
Mar 6, 2017·Biochemistry. Biokhimii︠a︡·A V ArtiukhovV I Bunik
Nov 20, 2020·International Journal of Molecular Sciences·Stefan R MarsdenUlf Hanefeld
Mar 2, 2021·AMB Express·Yajing LiangYi Jin
Sep 8, 2021·Nature Communications·Jana ŠkerlováPål Stenmark