Volume 14 Supplement 1

6th International Conference on cGMP: Generators, Effectors and Therapeutic Implications

Open Access

Mechanisms driving heme insertion into apo-sGC during its maturation in cells

BMC Pharmacology and Toxicology201314(Suppl 1):O18

DOI: 10.1186/2050-6511-14-S1-O18

Published: 29 August 2013

Background

During maturation of soluble guanylyl cyclase (sGC), heme insertion into the beta subunit is essential because it enables sGC to recognize nitric oxide (NO) and transduce its biological effects. We used a mammalian cell culture approach and followed heme insertion into both transiently- and endogenously-expressed apo-sGC beta. Although sGC is often associated with heat shock protein 90 (hsp90) in cells, the implications are unclear. Experiments that used pharmacological hsp90 inhibitors, an ATP-ase inactive hsp90 mutant, and heme-dependent or heme-independent sGC activators revealed that heme insertion into apo-sGC requires hsp90 [1].

Results

Our findings suggest a model where apo-sGC beta may directly complex with hsp90, which then drives heme insertion into apo-sGC beta through its inherent ATPase activity. In follow-up studies we are using purified proteins and domains and fluorescence and hydrogen-deuterium exchange methods to characterize the apo-sGC interaction with hsp90 at the molecular level. Results indicate that apo-sGC beta binds to the hsp90 middle domain, and the binding interaction involves structural regions within the heme and PAS domains of sGC.

Conclusion

Together, our work is shedding light on the sGC maturation process, and is revealing new ways that sGC activity could be impacted or controlled in cells.

Authors’ Affiliations

(1)
Department of Pathobiology, Lerner Research Institute, Cleveland Clinic

References

  1. Ghosh A, Stuehr DJ: Soluble guanylyl cyclase requires heat shock protein 90 for heme insertion during maturation of the NO-active enzyme. Proc Natl Acad Sci U S A. 2012, 109: 12998-13003. 10.1073/pnas.1205854109.PubMed CentralView ArticlePubMedGoogle Scholar

Copyright

© Stuehr et al; licensee BioMed Central Ltd. 2013

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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