title: Structural and functional analysis of betaine aldehyde dehydrogenase from Staphylococcus aureus.
abstract: When exposed to high osmolarity, methicillin-resistant Staphylococcus aureus (MRSA) restores its growth and establishes a new steady state by accumulating the osmoprotectant metabolite betaine. Effective osmoregulation has also been implicated in the acquirement of a profound antibiotic resistance by MRSA. Betaine can be obtained from the bacterial habitat or produced intracellularly from choline via the toxic betaine aldehyde (BA) employing the choline dehydrogenase and betaine aldehyde dehydrogenase (BADH) enzymes. Here, it is shown that the putative betaine aldehyde dehydrogenase SACOL2628 from the early MRSA isolate COL (SaBADH) utilizes betaine aldehyde as the primary substrate and nicotinamide adenine dinucleotide (NAD(+)) as the cofactor. Surface plasmon resonance experiments revealed that the affinity of NAD(+), NADH and BA for SaBADH is affected by temperature, pH and buffer composition. Five crystal structures of the wild type and three structures of the Gly234Ser mutant of SaBADH in the apo and holo forms provide details of the molecular mechanisms of activity and substrate specificity/inhibition of this enzyme.
authors.affiliation: Department of Biochemistry and Molecular Genetics, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
authors.forename: Andrei S
authors.initials: AS
authors.lastname: Halavaty
authors.affiliation: Biosensor Tools LLC, Salt Lake City, UT 84103, USA.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Rich
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, Ontario M5S 3E5, Canada.
authors.forename: Chao
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, Ontario M5S 3E5, Canada.
authors.forename: Jeong Chan
authors.initials: JC
authors.lastname: Joo
authors.affiliation: Department of Biochemistry and Molecular Genetics, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
authors.forename: George
authors.initials: G
authors.lastname: Minasov
authors.affiliation: Department of Biochemistry and Molecular Genetics, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
authors.forename: Ievgeniia
authors.initials: I
authors.lastname: Dubrovska
authors.affiliation: Department of Biochemistry and Molecular Genetics, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
authors.forename: James R
authors.initials: JR
authors.lastname: Winsor
authors.affiliation: Biosensor Tools LLC, Salt Lake City, UT 84103, USA.
authors.forename: David G
authors.initials: DG
authors.lastname: Myszka
authors.affiliation: Center for Structural Genomics of Infectious Diseases (CSGID), Chicago, IL 60611, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Duban
authors.affiliation: Department of Biochemistry and Molecular Genetics, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
authors.forename: Ludmilla
authors.initials: L
authors.lastname: Shuvalova
authors.affiliation: Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, Ontario M5S 3E5, Canada.
authors.forename: Alexander F
authors.initials: AF
authors.lastname: Yakunin
authors.affiliation: Department of Biochemistry and Molecular Genetics, Northwestern University, 303 East Chicago Avenue, Chicago, IL 60611, USA.
authors.forename: Wayne F
authors.initials: WF
authors.lastname: Anderson
mesh_terms: D001622:Betaine / Q000031:analogs & derivatives* / Q000378:metabolism; D050728:Betaine-Aldehyde Dehydrogenase / Q000737:chemistry* / Q000378:metabolism*; D001665:Binding Sites; D018360:Crystallography, X-Ray; D007700:Kinetics; D008958:Models, Molecular; D009243:NAD / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013211:Staphylococcus aureus / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D009243:NAD; D001622:Betaine; C026820:betaine aldehyde; D050728:Betaine-Aldehyde Dehydrogenase
keywords: Staphylococcus aureus; betaine aldehyde dehydrogenase; high-throughput approach; infectious diseases; structural genomics