Structure and activity of the NAD(P)+-dependent succinate semialdehyde dehydrogenase YneI from Salmonella typhimurium

Hongyan Zheng, Alina Beliavsky, Anatoli Tchigvintsev, Joseph S Brunzelle, Greg Brown, Robert Flick, Elena Evdokimova, Zdzislaw Wawrzak, Radhakrishnan Mahadevan, Wayne F Anderson, Alexei Savchenko, Alexander F. Yakunin*

*Corresponding author for this work

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Aldehyde dehydrogenases are found in all organisms and play an important role in the metabolic conversion and detoxification of endogenous and exogenous aldehydes. Genomes of many organisms including Escherichia coli and Salmonella typhimurium encode two succinate semialdehyde dehydrogenases with low sequence similarity and different cofactor preference (YneI and GabD). Here, we present the crystal structure and biochemical characterization of the NAD(P)+-dependent succinate semialdehyde dehydrogenase YneI from S. typhimurium. This enzyme shows high activity and affinity toward succinate semialdehyde and exhibits substrate inhibition at concentrations of SSA higher than 0.1 mM. YneI can use both NAD+ and NADP+ as cofactors, although affinity to NAD+ is 10 times higher. High resolution crystal structures of YneI were solved in a free state (1.85 Å) and in complex with NAD+ (1.90 Å) revealing a two domain protein with the active site located in the interdomain interface. The NAD+ molecule is bound in the long channel with its nicotinamide ring positioned close to the side chain of the catalytic Cys268. Site-directed mutagenesis demonstrated that this residue, as well as the conserved Trp136, Glu365, and Asp426 are important for activity of YneI, and that the conserved Lys160 contributes to the enzyme preference to NAD+. Our work has provided further insight into the molecular mechanisms of substrate selectivity and activity of succinate semialdehyde dehydrogenases.

Original languageEnglish (US)
Pages (from-to)1031-1041
Number of pages11
JournalProteins: Structure, Function and Bioinformatics
Volume81
Issue number6
DOIs
StatePublished - Jun 1 2013

Keywords

  • Aldehyde dehydrogenase
  • Crystal structure
  • Kinetics
  • Site-directed mutagenesis
  • Succinate semialdehyde

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology

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