Physical and Functional Interaction Between Calcineurin and the Cardiac L-Type Calcium Channel
dc.contributor.advisor | Hill, Joseph A. | en |
dc.creator | Tandan, Samvit | en |
dc.date.accessioned | 2010-07-12T18:15:52Z | |
dc.date.available | 2010-07-12T18:15:52Z | |
dc.date.issued | 2008-09-19 | |
dc.description.abstract | The L-type Ca2+ channel is the major mediator of Ca2+ influx in cardiomyocytes leading to both mechanical contraction and activation of signaling cascades. Among these cascades is calcineurin, a protein phosphatase that promotes hypertrophic growth of the heart. We previously reported from in vivo models of pressure-overload that calcineurin regulates Ca2+ channel function in the heart, such that, inhibition of calcineurin activity results in a decrease in channel function. Based on this, we hypothesize that calcineurin participates in the channel macromolecular complex. Initial immunohistochemical data demonstrated evidence for co-localization of calcineurin to alpha1C, the pore-forming subunit of the L-type Ca2+ channel in mouse ventricular tissue. Additionally, co-immunoprecipitation biochemical experiments revealed evidence for binding between calcineurin and alpha1C in native mouse and rat heart tissues. Pulldown assays using GST-fusion proteins of all intracellular alpha1C regions provided evidence for direct binding of calcineurin at the N- and C-termini of alpha1C. At the C-terminus, calcineurin bound to aa 1909-2029 overlapping the well-characterized PKA/PKC site Ser-1928. In vitro kinase/phosphatase assays revealed Ser-1928 as a substrate for calcineurin dephosphorylation. Voltage-clamp recordings of L-type Ca2+ currents from cultured cardiomyocytes expressing constitutively-active calcineurin revealed significant up-regulation of channel function, similar to our previous observations from cardiac hypertrophy in vivo. Conversely, acute suppression of calcineurin, both pharmacologically or with specific peptide-inhibitors, induced a significant decrease in L-type channel function, while neither intervention had an effect on channel function in the absence of calcineurin activity. These data provide evidence for direct interaction between the L-type Ca2+ channel and calcineurin, and insights into the regulation of the channel by calcineurin. Furthermore, they highlight the specific role of calcineurin as a potential mediator of pathophysiological electrical remodeling in cardiac hypertrophy and failure. | en |
dc.format.digitalOrigin | born digital | en |
dc.format.medium | Electronic | en |
dc.format.mimetype | application/pdf | en |
dc.identifier.oclc | 759168774 | |
dc.identifier.uri | https://hdl.handle.net/2152.5/491 | |
dc.language.iso | en | en |
dc.subject | Calcineurin | en |
dc.subject | Calcium Channels, L-Type | en |
dc.subject | Myocytes, Cardiac | en |
dc.title | Physical and Functional Interaction Between Calcineurin and the Cardiac L-Type Calcium Channel | en |
dc.type | Thesis | en |
dc.type.genre | dissertation | en |
dc.type.material | Text | en |
thesis.date.available | 2009-09-19 | |
thesis.degree.department | Graduate School of Biomedical Sciences | en |
thesis.degree.discipline | Integrative Biology | en |
thesis.degree.grantor | UT Southwestern Medical Center | en |
thesis.degree.level | Doctoral | en |
thesis.degree.name | Doctor of Philosophy | en |