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Title: Structural insights into the mechanism of oxidative activation of heme-free H-NOX from Vibrio cholerae

Journal Article · · Biochemical Journal
 [1];  [2];  [3];  [1]; ORCiD logo [1]
  1. New Mexico State Univ., Las Cruces, NM (United States). Dept. of Chemistry and Biochemistry
  2. Reed College, Portland, OR (United States). Dept. of Chemistry
  3. New Mexico State Univ., Las Cruces, NM (United States). Dept. of Plant and Environmental Sciences

Bacterial heme nitric oxide/oxygen (H-NOX) domains are nitric oxide (NO) or oxygen sensors. This activity is mediated through binding of the ligand to a heme cofactor. However, H-NOX from Vibrio cholerae (Vc H-NOX) can be easily purified in a heme-free state that is capable of reversibly responding to oxidation, suggesting a heme-independent function as a redox sensor. This occurs by oxidation of Cys residues at a zinc-binding site conserved in a subset of H-NOX homologs. Remarkably, zinc is not lost from the protein upon oxidation, although its ligation environment is significantly altered. Using a combination of computational and experimental approaches, we have characterized localized structural changes that accompany the formation of specific disulfide bonds between Cys residues upon oxidation. Furthermore, the larger-scale structural changes accompanying oxidation appear to mimic those changes observed upon NO binding to the heme-bound form. Thus, Vc H-NOX and its homologs may act as both redox and NO sensors by completely separate mechanisms.

Research Organization:
SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
AC02-76SF00515
OSTI ID:
1624989
Journal Information:
Biochemical Journal, Vol. 477, Issue 6; ISSN 0264-6021
Publisher:
Biochemical SocietyCopyright Statement
Country of Publication:
United States
Language:
English

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Figures / Tables (8)