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Title: Structural and functional analysis of betaine aldehyde dehydrogenase from Staphylococcus aureus

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. Finally, 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:
 [1] ;  [2] ;  [3] ;  [3] ;  [1] ;  [1] ;  [1] ;  [2] ;  [4] ;  [1] ;  [3] ;  [1]
  1. Northwestern Univ., Chicago, IL (United States). Dept. of Biochemistry and Molecular Genetics; Center for Structural Genomics of Infectious Diseases (CSGID), Chicago, IL (United States)
  2. Biosensor Tools LLC, Salt Lake City, UT (United States)
  3. Univ. of Toronto, ON (Canada). Dept. of Chemical Engineering and Applied Chemistry
  4. Center for Structural Genomics of Infectious Diseases (CSGID), Chicago, IL (United States)
Publication Date:
OSTI Identifier:
1233329
Grant/Contract Number:
AC02-06CH11357; 085P1000817; HHSN272200700058C; HHSN272201200026C
Type:
Accepted Manuscript
Journal Name:
Acta Crystallographica. Section D: Biological Crystallography (Online)
Additional Journal Information:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography (Online); Journal Volume: 71; Journal Issue: 5; Journal ID: ISSN 1399-0047
Publisher:
International Union of Crystallography
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org:
National Institutes of Health (NIH); USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; betaine aldehyde dehydrogenase; Staphylococcus aureus; structural genomics; high-throughput approach; infectious diseases