Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8289290 | Archives of Biochemistry and Biophysics | 2016 | 25 Pages |
Abstract
A phospho-null Ala substitution at protein kinase C (PKC)-targeted cardiac troponin I (cTnI) S43/45 reduces myocyte and cardiac contractile function. The goal of the current study was to test whether cTnIS43/45N is an alternative, functionally conservative substitution in cardiac myocytes. Partial and more extensive endogenous cTnI replacement was similar at 2 and 4 days after gene transfer, respectively, for epitope-tagged cTnI and cTnIS43/45N. This replacement did not significantly change thin filament stoichiometry. In functional studies, there were no significant changes in the amplitude and/or rates of contractile shortening and re-lengthening after this partial (2 days) and extensive (4 days) replacement with cTnIS43/45N. The cTnIS43/45N substitution also was not associated with adaptive changes in the myocyte Ca2+ transient or in phosphorylation of the protein kinase A and C-targeted cTnIS23/24 site. These results provide evidence that cTnIS43/45N is a functionally conservative substitution, and may be appropriate for use as a phospho-null in rodent models designed for studies on PKC modulation of cardiac performance.
Keywords
PKCGARGAMHRPcTnINGSABSFITCFBSDMEMECLPBSTX-100myofilamentDulbecco's modified Eagle MediumP/STnIantibodiescTnCgoat anti-rabbitenhanced chemiluminescenceanalysis of varianceANOVATropomyosinTroponincardiac troponin CTroponin ITriton X-100Texas redfetal bovine serumnormal goat serumgoat anti-mousePhosphate buffered salinePhosphorylationfluorescein isothiocyanateHeartcardiac troponin ISilver stainAntibodyHorseradish peroxidaseProtein kinase CPenicillin/streptomycin
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Authors
Sarah E. Lang, Tamara K. Stevenson, Dongyang Xu, Ryan O'Connell, Margaret V. Westfall,