Activation of endogenous protein phosphatase 1 enhances the calcium sensitivity of the ryanodine receptor type 2 in murine ventricular cardiomyocytes.
KEY POINTS Increased protein phosphatase 1 (PP-1) activity was found in the late stages of human heart failure. Although PP-1 has been studied extensively, a detailed understanding of its role in excitation-contraction coupling mechanism, in normal and diseased heart, remains elusive. This work examines the functional effects of the activities of PP-1 on the release of Ca2 + local events in ventricular cardiomyocytes, using the activation peptide (PDP3) for the stimulation of endogenous PP-1 protein. We reported that the acute de-phosphorylation may increase the sensitivity of RyR2 to Ca2 + channels in situ, and that RyR2-serine2808 phosphorylation sites can mediate the process. Our approach unmask the functional importance of PP-1 in the regulation of RyR2 activity, suggesting a potential role in the generation of the pathophysiology of the sarcoplasmic reticulum Ca2 + leak at the heart sick.
ABSTRACT Changes heart Ryanodine receptor (RyR2) phosphorylation was considered a disease of regulation and post-translational modification of proteins related matters. The extent of phosphorylation of RyR2 is mainly determined by the balance of the activity of protein kinases and phosphatases, respectively. Increased protein phosphatase-1 (PP-1) activity has been observed in heart failure, but the role of this enzyme in the regulation of intracellular Ca2 + handling the remains poorly understood.
To determine the physiological and significance of the pathophysiology of increased activity of PP-1, we investigate how the PP-1 catalytic subunit (PP-1c) alter Ca2 + sparks in cardiomyocytes permeabilized and we also apply PP-1-disturbing peptide (PDP3) to specifically activate endogenous PP-1, including one anchored in the RyR2 macromolecular complex.
We compared the wild type (WT) and transgenic mice in which the normally highly phosphorylated RyR2 site-S2808 has been ablated to investigate his involvement in RyR2 modulation (S2808A + / +). In WT myocytes, PP-1 increased the frequency of Ca2 + spark (CaSpF) by 2-fold, followed by depletion of sarcoplasmic reticulum (SR) Ca2 + stores. Similarly, PDP3 temporarily increase and decrease the frequency spark SR Ca2 + load. RyR2 Ca2 + sensitivity, as assessed by analysis of Ca2 + spark recovery, increase with PDP3 when compared to the negative control peptide. S2808A + / + cardiomyocytes do not respond well PP-1c and treatment PDP3.
Our results showed an increase in Ca 2+ sensitivity of RyR2 in the de-phosphorylation by PP-1. Furthermore, we confirmed the S2808 as a target site for PP-1 and as a potential relationship between the modulation RyR2s and cellular responses. This article is protected by copyright. All rights reserved.
Phactr one set Slack (KCNT one ) channels through protein phosphatase one (PP one ).
The Slack (KCNT1) gene encodes sodium-activated potassium channels are abundant expressed in the central nervous system. human mutations alter channel function Slack, resulting in epilepsy and intellectual disability. The majority of disease-causing mutations located in the cytoplasmic C-terminus extended channels and the result is increased current Slack Slack. Previous experiments have shown that the C-terminus of the protein channel Slack bind a number of cytoplasmic signaling.
Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC
Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is Unconjugated. Tested in the following application: ELISA, WB; Recommended dilution: WB:1:500-1:2000
Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is HRP conjugated. Tested in the following application: ELISA
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.
Description: Calcineurin is an enzyme that dephosphorylates serine and threonine residues in proteins. It is a heterodimer of a 59kDa catalytic A subunit and a 19kDa regulatory B subunit that is activated by the binding of calcium ions and calmodulin. Calcineurin is expressed in many tissues, but its levels are highest in the brain, where it may play a role in learning and memory. It has many substrates, including NFAT, a transcription factor that is activated by dephosphorylation. Complexes of the immuno-suppressants cyclosporine and FK506 with immunophilin proteins such as cyclophilin and FKBP12 are potent and specific inhibitors of Calcineurin activity. Alterations in Calcineurin activity are suspected to play a role in cardiac hypertrophy and graft versus host disease in organ transplantation.
Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA
Description: A polyclonal antibody against PPP3R1. Recognizes PPP3R1 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA
Description: Description of target: regulatory subunit of the enzyme that catalyzes the calcium-dependent removal of serine- and threonine-bound phosphate groups [RGD, Feb 2006];Species reactivity: Rat;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.086 ng/mL
PPP3R1 (Myc-DDK-tagged)-Human protein phosphatase 3, regulatory subunit B, alpha (PPP3R1)
Description: Human PPP3R1 knockdown cell line is engineered by our optimized transduction of the specific shRNA with lentivirus. Knockdown levels are determined via qRT-PCR. Gentaur offers generation of stable knockdown (RNAi) cell lines expressing shRNAs targeting genes of your interest.
One is Phactr1, actin-binding protein that recruits protein phosphatase 1 (PP1) to specific phosphoprotein substrates. Using co-immunoprecipitation, we found that Phactr1 required to connect the channel to actin. Using a clamp recordings, we found that co-expression of wild-type channels Phactr1 with Slack reduce current amplitude but had no effect on the channel Slack where preserved PKC phosphorylation sites (S407), which regulates the current amplitude has been mutated.