4MPB

1.7 Angstrom resolution crystal structure of betaine aldehyde dehydrogenase (betB) from Staphylococcus aureus

Replaces:  3ED6

Summary for 4MPB

• Entry DOI: 10.2210/pdb4mpb/pdb
• Related: 4MPY
• Descriptor: Betaine aldehyde dehydrogenase, MAGNESIUM ION, CHLORIDE ION, ... (4 entities in total)
• Functional Keywords: structural genomics, nad, center for structural genomics of infectious, niaid, national institute of allergy and infectious diseases, csgid, rossmann fold, oxidoreductase, center for structural genomics of infectious diseases
• Biological source: Staphylococcus aureus subsp. aureus
• Total number of polymer chains: 2
• Total formula weight: 115166.14

Authors

Halavaty, A.S.,Shuvalova, L.,Minasov, G.,Dubrovska, I.,Winsor, J.,Peterson, S.N.,Anderson, W.F.,Center for Structural Genomics of Infectious Diseases (CSGID) (deposition date: 2013-09-12, release date: 2013-09-25, Last modification date: 2024-11-06)

Primary citation

Chen, C.,Joo, J.C.,Brown, G.,Stolnikova, E.,Halavaty, A.S.,Savchenko, A.,Anderson, W.F.,Yakunin, A.F.
Structure-based mutational studies of substrate inhibition of betaine aldehyde dehydrogenase BetB from Staphylococcus aureus.
Appl.Environ.Microbiol., 80:3992-4002, 2014

PubMed Abstract: Inhibition of enzyme activity by high concentrations of substrate and/or cofactor is a general phenomenon demonstrated in many enzymes, including aldehyde dehydrogenases. Here we show that the uncharacterized protein BetB (SA2613) from Staphylococcus aureus is a highly specific betaine aldehyde dehydrogenase, which exhibits substrate inhibition at concentrations of betaine aldehyde as low as 0.15 mM. In contrast, the aldehyde dehydrogenase YdcW from Escherichia coli, which is also active against betaine aldehyde, shows no inhibition by this substrate. Using the crystal structures of BetB and YdcW, we performed a structure-based mutational analysis of BetB and introduced the YdcW residues into the BetB active site. From a total of 32 mutations, those in five residues located in the substrate binding pocket (Val288, Ser290, His448, Tyr450, and Trp456) greatly reduced the substrate inhibition of BetB, whereas the double mutant protein H448F/Y450L demonstrated a complete loss of substrate inhibition. Substrate inhibition was also reduced by mutations of the semiconserved Gly234 (to Ser, Thr, or Ala) located in the BetB NAD(+) binding site, suggesting some cooperativity between the cofactor and substrate binding sites. Substrate docking analysis of the BetB and YdcW active sites revealed that the wild-type BetB can bind betaine aldehyde in both productive and nonproductive conformations, whereas only the productive binding mode can be modeled in the active sites of YdcW and the BetB mutant proteins with reduced substrate inhibition. Thus, our results suggest that the molecular mechanism of substrate inhibition of BetB is associated with the nonproductive binding of betaine aldehyde.

PubMed: 24747910
DOI: 10.1128/AEM.00215-14

Experimental method

X-RAY DIFFRACTION (1.7 Å)