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Mass Spectrometry as a Novel Approach to Probe Cooperativity in Multimeric Enzymatic Systems by Hélène Rogniaux; Sarah Sanglier; Kerstin Strupat; Saı̈d Azza; Olivier Roitel; Vincent Ball; Denis Tritsch; Guy Branlant; Alain Van Dorsselaer is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Mass Spectrometry as a Novel Approach to Probe Cooperativity in Multimeric Enzymatic Systems about?

Investigating cooperativity in multimeric enzymes is of utmost interest to improve our understanding of the mechanism of enzymatic regulation. In the present article, we propose a novel approach based on mass spectrometry to probe cooperativity in the binding of a ligand to a multisubunit enzyme. This approach presents the selective advantage of giving a direct insight into all the subsequent ligation states that are formed in solution as the ligand is added to the enzyme. A quantitative interpretation of the electrospray ionization (ESI) mass spectra gives the relative abundance of all the distinct enzymatic species, which allows one to directly deduce the cooperativity of the system. The overall method is described for the addition of the oxidized cofactor nicotinamide adenine dinucleotide (NAD(+)) to a dimeric mutant of Bacillus stearothermophilus glyceraldehyde-3-phosphate dehydrogenase (GPDH). It is then applied to four tetrameric enzymes: sturgeon muscle GPDH, wild type and S48G mutant of GPDH from B. stearothermophilus, and alcohol dehydrogenase (ADH) from Bakers yeast. The results illustrate the possibilities offered by this new technique. First, mass spectrometry allows a

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Author

Hélène Rogniaux; Sarah Sanglier; Kerstin Strupat; Saı̈d Azza; Olivier Roitel; Vincent Ball; Denis Tritsch; Guy Branlant; Alain Van Dorsselaer

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)

Published

2001

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)