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Soluble Guanylate Cyclase
People working on this project: Emily Derbyshire
Shirley Huang
Michael Winter

The nitric oxide (NO) signaling pathway is important for many physiological functions including vascular smooth muscle relaxation, neuronal signal transduction and inhibition of platelet aggregation.  The source of NO in vivo is the enzyme nitric oxide synthase.  The principal receptor for NO is soluble guanylate cyclase (sGC), which catalyzes the conversion of GTP to the second messenger molecule cyclic GMP (cGMP).  sGC is a member of a family of related enzymes which share homologous catalytic domains, but are activated in different ways.  This family includes the adenylate cyclases, a class of membrane bound enzymes that convert ATP to cAMP and are regulated by G proteins, and the membrane-bound guanylate cyclases that make cGMP in response to hormone signals via an extracellular ligand binding domain.

sGC is a heterodimeric hemoprotein consisting of homologous alpha and beta subunits.  Each subunit consists of a N-terminal H-NOX domain, a central domain of unknown function, and a C-terminal consensus nucleotide cyclase domain.  When NO binds to the heme prosthetic group in the beta subunit of sGC, catalysis is accelerated by 2-3 orders of magnitude.

We are trying to answer a number of questions about sGC function and regulation, such as:
1. How does the binding of NO affect the heme and surrounding environment?
2. How is the NO binding event translated into increased catalytic activity?
3. How does sGC deactivate?
4. How do other small molecules such as carbon monoxide (CO), YC-1, and organic nitrogen oxides activate sGC?
5. How do nucleotides regulate sGC activity?

Additionally, we have initiated a series of experiments to characterize novel guanylate cyclases in C. elegans and Drosophila.  We have identified some of these putative cyclases through sequence analyses and cloned them, and are currently involved in both biochemical and in vivo characterization.
 

Selected sGC Publications from this laboratory:

  1. Winger, J.A., Derbyshire, E.R., and Marletta, M.A. (2007) Dissociation of nitric oxide from soluble guanylate cyclase and heme-nitric oxide/oxygen binding domain constructs. J. Biol. Chem. 282: 897-907.

  2. Cary, S.P.L., Winger, J.A., Derbyshire, E.R., and Marletta, M.A. (2006) Nitric oxide signaling: no longer simply on or off. TiBS 31: 231-239.

  3. Hering, K.W., Artz, J.D., Pearson, W.H., and Marletta, M.A. (2006) The design and synthesis of YC-1 analogues as probes for soluble guanylate cyclase. Bioorg. Med. Chem. Lett. 16: 618-621.

  4. Karow, D.S., Pan, D., Davis, J.H., Behrends, S., Mathies, R.A., and Marletta, M.A. (2005) Characterization of functional heme domains from soluble guanylate cyclase. Biochemistry 44: 16266-16274.

  5. Derbyshire, E.R. and Marletta, M.A. (2005) Characterization of nitrosoalkane binding and activation of soluble guanylate cyclase. Biochemistry 44: 16257-16265.

  6. Cary, S.P.L., Winger, J.A., and Marletta, M.A. (2005) Tonic and acute nitric oxide signaling through soluble guanylate cyclase is mediated by nonheme nitric oxide, ATP, and GTP. Proc. Natl. Acad. Sci. USA 102: 13064-13069.

  7. Winger, J.A. and Marletta, M.A. (2005) Expression and characterization of the catalytic domains of soluble guanylate cyclase: interaction with the heme domain. Biochemistry 44: 4083-4090.

  8. Gray, J.M., Karow, D.S., Lu, H., Chang, A.J., Chang, J.S., Ellis, R.E., Marletta, M.A., and Bargmann, C.I. (2004) Oxygen sensation and social feeding mediated by a C. elegans guanylate cyclase homologue. Nature 430: 317-22.

  9. Ballou, D. P., Zhao, Y., Brandish, P. E., and Marletta, M. A. (2002) Revisiting the kinetics of nitric oxide (NO) binding to soluble guanylate cyclase: the simple NO-binding model is incorrect. Proc. Natl. Acad. Sci. USA 99: 12097-12101.

  10. Artz, J.D., Schmidt, B., McCracken, J.L., and Marletta, M. A. (2002) Effects of Nitroglycerin on Soluble Guanylate Cyclase: Implications for Nitrate Tolerance. J. Biol. Chem. 277: 18253-18256.

  11. Zhao, Y., Brandish, P. E., DiValentin, M., Schelvis, J. P., Babcock, G. T. and Marletta, M. A. (2000) Inhibition of Soluble Guanylate Cyclase by ODQ. Biochemistry 39: 10848-10854.

  12. Zhao, Y., Brandish, P. E., Ballou, D. P. and Marletta, M. A. (1999) A molecular basis for nitric oxide sensing by soluble guanylate cyclase. Proc. Natl. Acad. Sci. USA 96: 14753-14758.

  13. Denninger, J.W. and Marletta, M.A. (1999) Guanylate cyclase and the NO/cGMP signaling pathway. Biochim. Biophys. Acta 1411: 334-350.

  14. Brandish, P. E., Buechler, W. and Marletta, M. A. (1998) Regeneration of the ferrous heme of soluble guanylate cyclase from the nitric oxide complex: acceleration by thiols and oxyhemoglobin. Biochemistry37: 16898-16907.

  15. Schelvis, J. P., Zhao, Y., Marletta, M. A. and Babcock, G. T. (1998) Resonance raman characterization of the heme domain of soluble guanylate cyclase. Biochemistry 37: 16289-16297.

  16. Zhao, Y., Hoganson, C., Babcock, G. T. and Marletta, M. A. (1998) Structural changes in the heme proximal pocket induced by nitric oxide binding to soluble guanylate cyclase. Biochemistry 37: 12458-12464.

  17. Stone, J.R. and Marletta, M.A. (1998) Synergistic activation of soluble guanylate cyclase by YC-1 and carbon monoxide: implications for the role of cleavage of the iron-histidine bond during activation by nitric oxide. Chem. Biol. 5: 255-261.

  18. Zhao, Y., Schelvis, J.P.M., Babcock, G.T. and Marletta, M.A. (1998) Identification of histidine 105 in the beta-1 subunit of soluble guanylate cyclase as the heme proximal ligand. Biochemistry 37: 4502-4509.

  19. Zhao, Y. and Marletta, M.A. (1997) Localization of heme binding region in soluble guanylate cyclase. Biochemistry 36: 15959-15964.

  20. Kim, S., Deinum, G., Gardner, M.T., Marletta, M.A. and Babcock, G.T. (1996) Distal pocket polarity in the unusual ligand binding site of soluble guanylate cyclase: implications for the control of NO binding. J. Am. Chem. Soc. 118: 8769-8770.

  21. Stone, J.R., Sands, R.H., Dunham, W.R. and Marletta, M.A. (1996) Spectral and ligand-binding properties of an unusual hemoprotein, the ferric form of soluble guanylate cyclase. Biochemistry 35: 3258-3262.

  22. Deinum, G., Stone, J.R., Babcock, G.T. and Marletta, M.A. (1996) Binding of nitric oxide and carbon monoxide to soluble guanylate cyclase as observed with resonance raman spectroscopy. Biochemistry35: 1540-1547.

  23. Stone, J.R. and Marletta, M.A. (1995) The ferrous heme of soluble guanylate cyclase: formation of hexacoordinate complexes with carbon monoxide and nitrosomethane. Biochemistry 34: 16397-16403.

  24. Stone, J.R., Sands, R.H., Dunham, W.R. and Marletta, M.A. (1995) Electron paramagnetic resonance spectral evidence for the formation of a pentacoordinate nitrosyl-heme complex on soluble guanylate cyclase. Biochem. Biophys. Res. Comm. 207: 572-577.

  25. Stone, J.R. and Marletta, M.A. (1994) Soluble guanylate cyclase from bovine lung: activation with nitric oxide and carbon monoxide and spectral characterization of the ferrous and ferric states. Biochemistry 33:5636-5640.
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