PMID: 7034777Dec 22, 1981Paper

Steady-state and pre-steady-state kinetics of coenzyme A linked aldehyde dehydrogenase from Escherichia coli

Biochemistry
C C Shone, H J Fromm

Abstract

Coenzyme A linked aldehyde dehydrogenase from Escherichia coli strain B has been purified to a specific activity of 14 units/mg of protein, and initial rate and substrate analogue inhibition experiments have been performed. On the basis of these steady-state measurements, a bi-uni-uni-uni ping-pong mechanism is proposed in which NAD+ binds to the free enzyme followed by acetaldehyde. The product NADH is then released before coenzyme A (CoA) can bind, and acetyl-CoA is the final product released. A pre-steady-state time-dependent activation of the enzyme was observed when assays were initiated with NAD+. This lag phase of the reaction was studied as a function of the NAD+ concentration and found to be first order. Furthermore, the presence of NAD+ was demonstrated to be necessary to maintain the enzyme in the active conformation. Evidence that the enzyme contains two distinct NAD+ binding sites, an activator site and a catalytic site, has been obtained from pre-steady-state experiments with the NAD+ analogues AMP and 3-pyridine-carboxaldehyde adenine dinucleotide. AMP, a potent competitive inhibitor with respect to NAD+ under steady-state conditions, did not affect the rate of enzyme activation during pre-steady-state measuremen...Continue Reading

Citations

Jul 21, 2009·Applied and Environmental Microbiology·Ashutosh GuptaRamon Gonzalez
Oct 13, 2000·Journal of Bacteriology·C A Holland-StaleyP R Cunningham
May 24, 2003·Proceedings of the National Academy of Sciences of the United States of America·Babu A ManjasettyAlice Vrielink
Apr 26, 2011·The Journal of Biological Chemistry·François TalfournierGuy Branlant
Jan 9, 2013·Chemico-biological Interactions·Lilian González-SeguraThomas D Hurley
Aug 23, 2011·Biotechnology and Bioengineering·Angela CintolesiRamon Gonzalez
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Mar 17, 2015·Proceedings of the National Academy of Sciences of the United States of America·Justin B SiegelDavid Baker
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