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dc.contributor.authorWeichsel, Andrzej
dc.contributor.authorKievenaar, Jessica A
dc.contributor.authorCurry, Roslyn
dc.contributor.authorCroft, Jacob T
dc.contributor.authorMontfort, William R
dc.date.accessioned2019-10-09T19:24:17Z
dc.date.available2019-10-09T19:24:17Z
dc.date.issued2019-10-01
dc.identifier.citationWeichsel A, Kievenaar JA, Curry R, Croft JT, Montfort WR. Instability in a coiled-coil signaling helix is conserved for signal transduction in soluble guanylyl cyclase. Protein Science. 2019; 28: 1830–1839. https://doi.org/10.1002/ pro.3707en_US
dc.identifier.issn0961-8368
dc.identifier.pmid31411784
dc.identifier.doi10.1002/pro.3707
dc.identifier.urihttp://hdl.handle.net/10150/634711
dc.description.abstractHow nitric oxide (NO) activates its primary receptor, α1/β1 soluble guanylyl cyclase (sGC or GC‐1), remains unknown. Likewise, how stimulatory compounds enhance sGC activity is poorly understood, hampering development of new treatments for cardiovascular disease. NO binding to ferrous heme near the N‐terminus in sGC activates cyclase activity near the C‐terminus, yielding cGMP production and physiological response. CO binding can also stimulate sGC, but only weakly in the absence of stimulatory small‐molecule compounds, which together lead to full activation. How ligand binding enhances catalysis, however, has yet to be discovered. Here, using a truncated version of sGC from Manduca sexta, we demonstrate that the central coiled‐coil domain, the most highly conserved region of the ~150,000 Da protein, not only provides stability to the heterodimer but is also conformationally active in signal transduction. Sequence conservation in the coiled coil includes the expected heptad‐repeating pattern for coiled‐coil motifs, but also invariant positions that disfavor coiled‐coil stability. Full‐length coiled coil dampens CO affinity for heme, while shortening of the coiled coil leads to enhanced CO binding. Introducing double mutation αE447L/βE377L, predicted to replace two destabilizing glutamates with leucines, lowers CO binding affinity while increasing overall protein stability. Likewise, introduction of a disulfide bond into the coiled coil results in reduced CO affinity. Taken together, we demonstrate that the heme domain is greatly influenced by coiled‐coil conformation, suggesting communication between heme and catalytic domains is through the coiled coil. Highly conserved structural imperfections in the coiled coil provide needed flexibility for signal transduction.en_US
dc.description.sponsorshipNational Cancer InstituteUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Cancer Institute (NCI) [P30 CA023074, U54 CA143924]; National Institute of General Medical SciencesUnited States Department of Health & Human ServicesNational Institutes of Health (NIH) - USANIH National Institute of General Medical Sciences (NIGMS) [R01 GM117357, R25 GM121228, T32 GM008804, T34 GM008718]; American Heart AssociationAmerican Heart Association [16PRE31090034]en_US
dc.language.isoenen_US
dc.publisherWILEYen_US
dc.rightsCopyright © 2019 The Protein Society.en_US
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/
dc.subjectGC-1en_US
dc.subjectYC-1en_US
dc.subjectcGMPen_US
dc.subjectguanylate cyclaseen_US
dc.subjectleucine zipperen_US
dc.subjectnitric oxideen_US
dc.subjectstimulator compounden_US
dc.titleInstability in a coiled-coil signaling helix is conserved for signal transduction in soluble guanylyl cyclaseen_US
dc.typeArticleen_US
dc.contributor.departmentUniv Arizona, Dept Chem & Biochemen_US
dc.identifier.journalPROTEIN SCIENCEen_US
dc.description.note12 month embargo; published online: 14 August 2019en_US
dc.description.collectioninformationThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.en_US
dc.eprint.versionFinal accepted manuscripten_US
dc.source.journaltitleProtein science : a publication of the Protein Society


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