title: Complex modulation of L-type Ca(2+) current inactivation by sorcin in isolated rabbit cardiomyocytes.
abstract: Modulation of the L-type Ca(2+) channel (LTCC) by sorcin was investigated by measuring the L-type Ca(2+) current (I (Ca,L)) in isolated rabbit ventricular myocytes using ruptured patch, single electrode voltage clamp in the absence of extracellular Na(+). Fifty millimolars EGTA (170 nM Ca(2+)) in the pipette solution buffered bulk cytoplasmic [Ca(2+)], but retained rapid Ca(2+)-dependant inactivation of I (Ca,L,). Recombinant sorcin (3 microM) in the pipette significantly slowed time-dependant inactivation (tau (fast): 8.8 +/- 0.9 vs. 15.1 +/- 1.7 ms). Sorcin had no significant effect on I (Ca,L,) after inhibition of the sarcoplasmic reticulum (SR). Using 10 mM 1,2-bis(o-N,N,N',N'-tetraacetic acid (170 nM Ca(2+)), I (Ca,L) inactivation was then determined by a Ca(2+) -independent, voltage-dependant process. Under these conditions, 3 microM sorcin speeded up inactivation. A similar effect was observed by substitution of Ca(2+) with Ba(2+). Down-regulation of endogenous sorcin to 27 +/- 7% using an RNAi adenoviral vector slowed inactivation of I (Ca,L) by approximately 42%. The effects of sorcin on voltage-dependant inactivation were mimicked by a truncated form of the protein containing only the Ca(2+)-binding domain. This data is consistent with two independent actions of sorcin on the LTCC: (1) slowing Ca(2+)-dependant inactivation and (2) stimulating voltage-dependant inactivation. The net effect of sorcin on the time-dependent inactivation of I (Ca,L) was a balance between these two effects. Under normal conditions, sorcin slows I (Ca,L) inactivation because the effects of Ca(2+)-dependant inactivation out-weigh the effects on voltage-dependant inactivation.
authors.affiliation: University of Glasgow, UK.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Fowler
authors.forename: Gianni
authors.initials: G
authors.lastname: Colotti
authors.forename: Emilia
authors.initials: E
authors.lastname: Chiancone
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Higuchi
authors.forename: Tim
authors.initials: T
authors.lastname: Seidler
authors.forename: Godfrey L
authors.initials: GL
authors.lastname: Smith
mesh_terms: D000818:Animals; D020746:Calcium Channels, L-Type / Q000187:drug effects / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D032383:Myocytes, Cardiac / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D034622:RNA Interference; D011817:Rabbits
chemical_list: D020746:Calcium Channels, L-Type; D002135:Calcium-Binding Proteins; D047428:Protein Kinase Inhibitors; D004533:Egtazic Acid; D017868:Cyclic AMP-Dependent Protein Kinases; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid