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The Marletta Laboratory

Bacterial Gas Sensing

People working on this project:
Mark Herzik
Charles Hespen
Alexander Nierth, PhD
Lars Plate

Heme-Nitric Oxide/Oxygen (H-NOX) proteins are a family of gas sensing proteins that are prevalent in prokaryotes and higher eukaryotes. H-NOX proteins were initially identified as homologs of the heme domain of the mammalian nitric oxide receptor soluble guanylate cyclase (sGC).

Biological function of H-NOX proteins

Genomic analysis reveals that prokaryotic H-NOX domains are linked to histidine kinases, diguanylate cyclases, and methyl-accepting chemotaxis proteins, suggesting involvement in prokaryotic signaling processes. Uncovering the biological function of these H-NOX domains, in particular in pathogens, is currently an area of intense investigation in our lab.

HNOX HK c-di-GMP

Ligand affinity and selectivity

H-NOX proteins exhibit remarkable diatomic ligand selectivity: Proteins from facultative aerobes exclusively bind NO, like sGC, whereas the H-NOX domains of most obligate anaerobes bind NO and O2. Structural and biochemical studies have shed light on the mechanism of ligand selectivity and highlighted the importance of a hydrogen-bonding network in the distal heme pocket to stabilize oxygen as a ligand. Our lab uses a variety of spectroscopic, kinetic, and structural techniques to probe how changes in protein conformation, dynamics, heme distortion, and protein tunnels affect ligand affinity and selectivity.

Ligand affinity and selectivity research image

Molecular mechanism of H-NOX signal transduction

Prokaryotic H-NOX domains are sensors for diatomic gases and as such transfer this sensory input to another signaling partner in the cell. Our lab is investigating the conformational changes that occur when ligands bind to the H-NOX protein and how these are communicated to partner proteins (histidine kinases, diguanylate cyclases, or methyl accepting chemotaxis proteins) to initiate the cascade of signaling events in the cell.

Ligand affinity and selectivity research image

H-NOX tool development

The H-NOX protein from Thermoanaerobacter tengcongensis is a small, readily expressed, remarkably stable protein that can accept a variety of porphyrin analogs. These characteristics inspired our interest in using this porphyrin-binding protein as a biological platform for the development of novel biotechnological tools. One such application was to create an O2-sensor by substituting the natural iron porphyrin with a ruthenium analog, which exhibits O2-dependent phosphorescence. Currently, we are working on utilizing H-NOX proteins to design new tools for medical or biotechnological applications.

H-NOX tool development research image

Publications

Plate L, Marletta MA. Nitric oxide modulates bacterial biofilm formation through a multicomponent cyclic-di-GMP signaling network. Mol. Cell. 2012 46:449-60.

Winter MB, Herzik MA Jr, Kuriyan J, Marletta MA. Tunnels modulate ligand flux in a heme nitric oxide/oxygen binding (H-NOX) domain. Proc. Natl. Acad. Sci. USA. 2011 108: E881-9.

Weinert EE, Phillips-Piro CM, Tran R, Mathies RA, Marletta MA. Controlling Conformational Flexibility of an O2-binding N-NOX Domain. Biochemistry. 2011 50(32):6832-40.

Tran R, Weinert EE, Boon EM, Mathies RA, Marletta MA. Determinants of the Heme-CO Vibrational Modes in the H-NOX Family. Biochemistry. 2011 50(30): 6519-30.

Olea C, Kuriyan J, Marletta MA. Modulating heme redox potential through protein-induced porphyrin distortion. J Am Chem Soc. 2010, 132: 12794-5.

Carlson HK, Vance RE, Marletta MA. H-NOX regulation of c-di-GMP Metabolism and Biofilm Formation in Legionella pneumophila. Mol Microbiol 2010, 77(4):930-42.

Wang Y, Dufour YS, Carlson HK, Donohue TJ, Marletta MA, Ruby EG. H-NOX-mediated nitric oxide sensing modulates symbiotic colonization by Vibrio fischeri. Proc Natl Acad Sci USA. 2010, 107: 8375-80.

Winter MB, McLaurin EJ, Reece SY, Olea C, Nocera DG, Marletta MA. Ru-Porphyrin Protein Scaffolds for Sensing O2. J Am Chem Soc. 2010, 132, 5582-83.

Olea C Jr, Herzik MA Jr, Kuriyan J, Marletta MA. Structural insights into the molecular mechanism of H-NOX activation. Protein Sci. 2010, 19, 881-7.

Weinert EE, Plate L, Whited CA, Olea C Jr, Marletta MA. Determinants of Ligand Affinity and Heme Reactivity in H-NOX Domains. Angew Chem Int Ed Engl. 2010, 49, 720-23.

Erbil WK, Price MS, Wemmer DE, Marletta MA. A structural basis for H-NOX signaling in Shewanella oneidensis by trapping a histidine kinase inhibitory conformation. Proc Natl Acad Sci USA.2009, 106, 19753-60.

Carlson HK, Plate L, Price MS, Allen JJ, Shokat KM, Marletta MA. Use of a semisynthetic epitope to probe histidine kinase activity and regulation. Anal Biochem. 2010, 397, 139-43.

Tran R, Boon EM, Marletta MA, Mathies RA. Resonance Raman spectra of an O2-binding H-NOX domain reveal heme relaxation upon mutation. Biochemistry 2009, 48, 8568-77.

Olea C, Boon EM, Pellicena P, Kuriyan J, Marletta MA. Probing the Function of Heme Distortion in the H-NOX Family. ACS Chem. Biol. 2008, 3, 703-710.

Price MS, Chao LY, Marletta MA. Shewanella oneidensis MR-1 H-NOX Regulation of a Histidine Kinase by Nitric Oxide. Biochemistry 2007, 46, 13677-83.

Boon EM, Davis JH, Tran R, Karow DS, Huang SH, Pan D, Miazgowicz MM, Mathies RA, Marletta MA. Nitric oxide binding to prokaryotic homologs of the soluble guanylate cyclase beta1 H-NOX domain. J Biol Chem. 2006, 281, 21892-902.

Boon EM, Marletta MA. Sensitive and Selective Detection of Nitric Oxide Using an H-NOX Domain. J. Am. Chem. Soc. 2006, 128, 10022-3.

Boon EM, Marletta MA. Ligand discrimination in soluble guanylate cyclase and the H-NOX family of heme sensor proteins. Curr Opin Chem Biol. 2005, 9, 441-6.

Boon EM, Huang SH, Marletta MA. A molecular basis for NO selectivity in soluble guanylate cyclase. Nat Chem Biol. 2005, 1, 53-9

Boon EM, Marletta MA. Ligand specificity of H-NOX domains: from sGC to bacterial NO sensors. J Inorg Biochem. 2005, 99, 892-902.

Karow DS, Pan D, Tran R, Pellicena P, Presley A, Mathies RA, Marletta MA. Spectroscopic characterization of the soluble guanylate cyclase-like heme domains from Vibrio cholerae and Thermoanaerobacter tengcongensis. Biochemistry. 2004, 43, 10203-11.

Pellicena P, Karow DS, Boon EM, Marletta MA, Kuriyan J. Crystal structure of an oxygen-binding heme domain related to soluble guanylate cyclases. Proc. Natl. Acad. Sci. USA. 2004, 101, 12854-9.