Structural and Functional Evidence Indicates Selective Oxygen Signaling in Caldanaerobacter subterraneus H-NOX.
Acute and specific sensing of diatomic gas molecules is an essential facet of biological signaling. Heme nitric oxide/oxygen binding (H-NOX) proteins are a family of gas sensors found in diverse classes of bacteria and eukaryotes. The most commonly characterized bacterial H-NOX domains are from facultative anaerobes and are activated through a conformational change caused by formation of a 5-coordinate Fe(II)-NO complex. Members of this H-NOX subfamily do not bind O2 and therefore can selectively ligate NO even under aerobic conditions. In contrast, H-NOX domains encoded by obligate anaerobes do form stable 6-coordinate Fe(II)-O2 complexes by utilizing a conserved H-bonding network in the ligand-binding pocket. The biological function of O2-binding H-NOX domains has not been characterized. In this work, the crystal structures of an O2-binding H-NOX domain from the thermophilic obligate anaerobe Caldanaerobacter subterraneus (Cs H-NOX) in the Fe(II)-NO, Fe(II)-CO, and Fe(II)-unliganded states are reported. The Fe(II)-unliganded structure displays a conformational shift distinct from the NO-, CO-, and previously reported O2-coordinated structures. In orthogonal signaling assays using Cs H-NOX and the H-NOX signaling effector histidine kinase from Vibrio cholerae (Vc HnoK), Cs H-NOX regulates Vc HnoK in an O2-dependent manner and requires the H-bonding network to distinguish O2 from other ligands. The crystal structures of Fe(II) unliganded and NO- and CO-bound Cs H-NOX combined with functional assays herein provide the first evidence that H-NOX proteins from obligate anaerobes can serve as O2 sensors.
Department of Molecular and Cell Biology, University of California-Berkeley , 356 Stanley Hall, Berkeley, California 94720-3220, United States.
Charles W
CW
Hespen
QB3 Institute, University of California-Berkeley , 356 Stanley Hall, Berkeley, California, 94720-3220, United States.
Joel J
JJ
Bruegger
Department of Chemistry, HAC 416 Franklin & Marshall College , Lancaster, Pennsylvania 17604-3003, United States.
Christine M
CM
Phillips-Piro
Department of Molecular and Cell Biology, University of California-Berkeley , 356 Stanley Hall, Berkeley, California 94720-3220, United States.|Department of Chemistry Department of Molecular and Cell Biology QB3 Institute, University of California-Berkeley , 374B Stanley Hall, Berkeley, California 94720-3220, United States.
Michael A
MA
Marletta
D018360:Crystallography, X-Ray; D005296:Ferrous Compounds / Q000378:metabolism; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D011487:Protein Conformation; D015398:Signal Transduction; D046968:Thermoanaerobacter / Q000378:metabolism*
D005296:Ferrous Compounds; D009569:Nitric Oxide; D010100:Oxygen