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Probing domain interactions in soluble guanylate cyclase.

Publication ,  Journal Article
Derbyshire, ER; Winter, MB; Ibrahim, M; Deng, S; Spiro, TG; Marletta, MA
Published in: Biochemistry
May 2011

Eukaryotic nitric oxide (NO) signaling involves modulation of cyclic GMP (cGMP) levels through activation of the soluble isoform of guanylate cyclase (sGC). sGC is a heterodimeric hemoprotein that contains a Heme-Nitric oxide and OXygen binding (H-NOX) domain, a Per/ARNT/Sim (PAS) domain, a coiled-coil (CC) domain, and a catalytic domain. To evaluate the role of these domains in regulating the ligand binding properties of the heme cofactor of NO-sensitive sGC, we constructed chimeras by swapping the rat β1 H-NOX domain with the homologous region of H-NOX domain-containing proteins from Thermoanaerobacter tengcongensis, Vibrio cholerae, and Caenorhabditis elegans (TtTar4H, VCA0720, and Gcy-33, respectively). Characterization of ligand binding by electronic absorption and resonance Raman spectroscopy indicates that the other rat sGC domains influence the bacterial and worm H-NOX domains. Analysis of cGMP production in these proteins reveals that the chimeras containing bacterial H-NOX domains exhibit guanylate cyclase activity, but this activity is not influenced by gaseous ligand binding to the heme cofactor. The rat-worm chimera containing the atypical sGC Gcy-33 H-NOX domain was weakly activated by NO, CO, and O(2), suggesting that atypical guanylate cyclases and NO-sensitive guanylate cyclases have a common molecular mechanism for enzyme activation. To probe the influence of the other sGC domains on the mammalian sGC heme environment, we generated heme pocket mutants (Pro118Ala and Ile145Tyr) in the β1 H-NOX construct (residues 1-194), the β1 H-NOX-PAS-CC construct (residues 1-385), and the full-length α1β1 sGC heterodimer (β1 residues 1-619). Spectroscopic characterization of these proteins shows that interdomain communication modulates the coordination state of the heme-NO complex and the heme oxidation rate. Taken together, these findings have important implications for the allosteric mechanism of regulation within H-NOX domain-containing proteins.

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Published In

Biochemistry

DOI

EISSN

1520-4995

ISSN

0006-2960

Publication Date

May 2011

Volume

50

Issue

20

Start / End Page

4281 / 4290

Related Subject Headings

  • Vibrio cholerae
  • Thermoanaerobacter
  • Soluble Guanylyl Cyclase
  • Recombinant Fusion Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Rats
  • Protein Structure, Quaternary
  • Protein Multimerization
  • Oxygen
  • Nitric Oxide
 

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Derbyshire, E. R., Winter, M. B., Ibrahim, M., Deng, S., Spiro, T. G., & Marletta, M. A. (2011). Probing domain interactions in soluble guanylate cyclase. Biochemistry, 50(20), 4281–4290. https://doi.org/10.1021/bi200341b
Derbyshire, Emily R., Michael B. Winter, Mohammed Ibrahim, Sarah Deng, Thomas G. Spiro, and Michael A. Marletta. “Probing domain interactions in soluble guanylate cyclase.Biochemistry 50, no. 20 (May 2011): 4281–90. https://doi.org/10.1021/bi200341b.
Derbyshire ER, Winter MB, Ibrahim M, Deng S, Spiro TG, Marletta MA. Probing domain interactions in soluble guanylate cyclase. Biochemistry. 2011 May;50(20):4281–90.
Derbyshire, Emily R., et al. “Probing domain interactions in soluble guanylate cyclase.Biochemistry, vol. 50, no. 20, May 2011, pp. 4281–90. Epmc, doi:10.1021/bi200341b.
Derbyshire ER, Winter MB, Ibrahim M, Deng S, Spiro TG, Marletta MA. Probing domain interactions in soluble guanylate cyclase. Biochemistry. 2011 May;50(20):4281–4290.
Journal cover image

Published In

Biochemistry

DOI

EISSN

1520-4995

ISSN

0006-2960

Publication Date

May 2011

Volume

50

Issue

20

Start / End Page

4281 / 4290

Related Subject Headings

  • Vibrio cholerae
  • Thermoanaerobacter
  • Soluble Guanylyl Cyclase
  • Recombinant Fusion Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Rats
  • Protein Structure, Quaternary
  • Protein Multimerization
  • Oxygen
  • Nitric Oxide