GPMDB00575OtherReinartz M, et al.InstrumentNot AvailableNot availableMus_musculus_viruses, MouseMichael.Reinartz@uni-duesseldorf.de25162660Institut fuer Herz- und Kreislaufphysiologiehttp://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003500http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003337http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003647http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003525http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003380http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003443http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003586Not availableGPMDBThe protein kinase AKT is a central kinase in the heart and has a major impact on growth/hypertrophy, survival/apoptosis, and metabolism. To gain more insight into AKT isoform-specific signaling at the molecular level, we investigated the phosphoproteome of HL-1 cardiomyocytes carrying AKT1 or AKT2 isoform-specific knock down, respectively. We combined stable isotope labeling with high resolution mass spectrometry and identified 377 regulated phosphopeptides. Although AKT1 is expressed at 4-fold higher levels, insulin stimulation mainly activated AKT2, which might in part rely on a preferred interaction of AKT2 with the mammalian target of rapamycin complex 2. In line with this result, the highest number of regulated phosphopeptides was identified in the AKT2 knock down cells. Isoform-specific regulation of AKT targets not previously described could be observed, and specific regulation of indirect target sites allows a deeper insight into affected biological processes. In the myocardial context, we identified many phosphosites supporting a connection of AKT to excitation-contraction coupling. Phosphoproteins identified included L-type calcium channel, ryanodine receptor, junctophilin, histidine-rich calcium binding protein, phospholamban, heat shock protein beta-6, and Ca²⁺/calmodulin-dependent kinase II. In conclusion, AKT isoform-specific knock down combined with quantitative phosphoproteomics provided a powerful strategy to unravel AKT isoform-specific signaling.AKT1 and AKT2 induce distinct phosphorylation patterns in HL-1 cardiac myocytes.Reinartz Michael M,Raupach Annika A,Kaisers Wolfgang W,Gödecke Axel A,Reinartz Michael M, Raupach Annika A, Kaisers Wolfgang W, Gödecke Axel AHIHGHH, PRKBB, PRKBA, ASGP-R 1, distinct, HL-1, ASGPR 1, Hepatic lectin 1., 2410016A19Rik, Asgr-1, Phosphorylations, proto-oncogene c-Akt, RAC-PK-alpha, AKT1, Hepatic lectin H1, PKB|Akt, phosphorylation, PKBB, protein kinase B, AKT1 kinase, Asgr, ASGPR1, PKB, CWS6, PKBbeta, AKT, RAC-BETA, C-type lectin domain family 4 member H1, Akt, CLEC4H1, AW554154, PKBalpha, ASGPR, PKB-ALPHA, mHL-1, RAC, Rac, PKBBETA, RAC-ALPHA, thymoma viral proto-oncogeneSoluble, PRKBB, Insulin B Chain, PRKBA, biological signaling, Ass-1, FKBP12-rapamycin complex-associated protein, number, Insulin B, Spectrum Analyses, AKT1, Isotope-Coded Affinity, Hepatic lectin 1, C-type lectin domain family 4 member H1, CLEC4H1, AA408052, Insulin, Mass, fold, Cell., Carrying, Analysis, Isotopically-Coded Affinity, Mammalian target of rapamycin, Isotopically-Coded Affinity Tagging, Mass Spectroscopy, Mass Spectrum Analysis, Chain, Analyses, ASGPR 1, Asgr-1, number of, PKB|Akt, PKBB, ASS, ASGPR1, Iletin, Stable, AKT, Akt, signaling process, has or lacks parts of type, Isotope-Coded Affinity Tagging, Tagging, Stable Isotope Labeling, single organism signaling, FK506-binding protein 12-rapamycin complex-associated protein 1, Mechanistic target of rapamycin, data, Regular, HL-1, 2410016A19Rik, extra or missing physical or functional parts, RAC-PK-alpha, mTOR, Labeling, Spectrum Analysis, AKT1 kinase, Regular Insulin, Spectroscopy, mereological quality, Asgr, Isotope, 2.7.11.1, Experiment, PKB, CWS6, Sodium Insulin, PKBbeta, RAC-BETA, Stable Isotope, ASGPR, PKB-ALPHA, Insulin A Chain, mHL-1, Sodium, Novolin, PKBBETA, Soluble Insulin, Isotope Coded Affinity Tagging, Mass Spectrum Analyses, RAPT1, serine-threonine kinase Akt, Mass Spectrum, HIHGHH, ASGP-R 1, INS, Rapamycin and FKBP12 target 1, Spectrometry, Rapamycin target protein 1, proto-oncogene c-Akt, Isotope Labeling, Hepatic lectin H1, Affinity Tagging, Isotopically-Coded, signalling, protein kinase B, signalling process, AW554154, PKBalpha, cardinality, Isotope-Coded, RAC, Rac, Proteomes, RAC-ALPHA, thymoma viral proto-oncogeneHIHGHH, PRKBB, PRKBA, ASGP-R 1, distinct, HL-1, ASGPR 1, Hepatic lectin 1., 2410016A19Rik, Asgr-1, Phosphorylations, proto-oncogene c-Akt, RAC-PK-alpha, AKT1, Hepatic lectin H1, PKB|Akt, phosphorylation, PKBB, protein kinase B, AKT1 kinase, Asgr, ASGPR1, PKB, CWS6, PKBbeta, AKT, RAC-BETA, C-type lectin domain family 4 member H1, Akt, CLEC4H1, AW554154, PKBalpha, ASGPR, PKB-ALPHA, mHL-1, RAC, Rac, PKBBETA, RAC-ALPHA, thymoma viral proto-oncogenebiochemical pathways, projections, L Type VDCC alpha 1 Subunit, Soluble, PRKBB, Insulin B Chain, PRKBA, Metabolic Process, Herz, Phosphodiesterase Activator, CALML2, Calcium Dependent, Ass-1, Calcium 40, Metabolic Concepts, Receptors, Calcium-Dependent Activator Protein, junctophilin-1, adult heart, Cyclic AMP-Phosphodiesterase Activator, AKT1, type I programmed cell death, L-Histidine, Circulatory Collapse, circulatory vessel, rat, L-Type VDCC alpha-2 Subunit, Extrinsic Pathway Apoptoses, CLEC4H1, junctophilin-2, Hot, AA408052, junctophilin-3, Concepts, fold, Heat, L Type VDCC, Kinase, Analysis, Metabolism Concept, Phenomenon, L-type calcium channel, Isotopically-Coded Affinity Tagging, Mass Spectrum Analysis, Shock, CALM, calcio, Analyses, N-type calcium channel, heart or heart like organ, catabolism, L Type VDCC delta Subunit, Asgr-1, Cyclic AMP-Phosphodiesterase, proteins, metabolic process resulting in cell growth, number of, cardiac structure, PKB|Akt, ASS, Intrinsic Pathway Apoptosis, ASGPR1, Calcium Channels, caffeine-sensitive calcium-release channel, L-Type, 2-amino-3-(1H-imidazol-4-yl)propanoic acid, CAMC, signaling process, papilla, Calcium-Dependent Activator, biotransformation, ATP Phosphotransferases, Tagging, Catabolism, CAM3, Stable Isotope Labeling, ATP, single organism signaling, Blood Coagulation Factor IV, Ca, incorporation, vertebrate heart, dorsal tube, cellular suicide, Regular, anatomical protrusion, Coagulation, Process, P-type calcium channel, metabolism resulting in cell growth, L-Type VDCC delta Subunit, lamina, mouse, flanges, Calcium-Dependent Regulator, L Type VDCC beta Subunit, L-Type Calcium Channels, extra or missing physical or functional parts, Circulatory Failure, RAC-PK-alpha, Dihydropyridine, Spectrum Analysis, AKT1 kinase, Regular Insulin, Pln protein, Spectroscopy, Asgr, L Type Voltage Dependent Calcium Channels, chambered heart, signaling (initiator) caspase activity, induction of apoptosis, count, Intrinsic Pathway, Extrinsic Pathway Apoptosis, ASGPR, shelf, Activator Protein, cardium, Calcium Channel, Bovine Activator, Soluble Insulin, Phosphodiesterase Activating, Hypertrophies, Transphosphorylases, Long-Lasting, death rate, growth pattern, non-developmental growth, shelves, L-isomer, Spectrometry, proto-oncogene c-Akt, projection, ridge, turnover, human, Dihydropyridine Receptor, protein kinase B, L isomer, Intrinsic Pathway Apoptoses, induction of apoptosis by p53, spine, Phosphodiesterase Protein Activator, PLN protein, L-Type VDCC alpha-1 Subunit, Extrinsic Pathway, high voltage-dependent calcium channel activity, AMP-Phosphodiesterase Activator, Hot Temperatures, E430016J11Rik, Apoptoses, RAC-ALPHA, biological signaling, Entire heart, Processes, lamellae, Circulatory, caspase-dependent programmed cell death, number, Histidine, branchial heart, CAMIII, Insulin B, Spectrum Analyses, Phosphodiesterase Activating Factor, Calcium Dependent Regulator, Metabolic Processes, apoptotic programmed cell death, Isotope-Coded Affinity, ryanodine receptor, process of organ, presence, activation of apoptosis, Hepatic lectin 1, Failure, protrusion, lamella, Calcium Dependent Activator Protein, Dihydropyridine Receptors, C-type lectin domain family 4 member H1, mTOR complex, 20Ca, Metabolism, Insulin, Factor IV, Mass, Carrying, L Type, Protein Activator, Type I, Isotopically-Coded Affinity, Metabolism Phenomena, Mass Spectroscopy, Bovine Activator Protein, Cyclic AMP Phosphodiesterase Activator, Chain, Calcium-40, L Type VDCC alpha 2 Subunit, ASGPR 1, Long Lasting Calcium Channels, ligand, Metabolic Concept, Activator, ridges, apoptosis activator activity, L-Type VDCC gamma Subunit, Activating Factor, PKBB, Hypovolemic Shock, Phosphodiesterase Protein, Hypovolemic, cardiac pump, Iletin, Bovine, Plm protein, Stable, AKT, Akt, time of survival, execution phase of apoptotic process, has or lacks parts of type, Isotope-Coded Affinity Tagging, programmed cell death by apoptosis, laminae, single organism signaling., histidine, Apoptosis, cell suicide, HL-1, L-isomer Histidine, degradation, PLB protein, anatomical process, CaM, apoptotic cell death, 2410016A19Rik, phospholamban protein, Hearts, Factor, Kalzium, Labeling, Phosphotransferases, Cell, L Type Calcium Channel, Concept, Metabolic Phenomena, mereological quality, polypeptide, Metabolism Concepts, Isotope, count in organism, PKB, VDCC, survival, CWS6, Sodium Insulin, Temperatures, PKBbeta, RAC-BETA, Stable Isotope, Programmed Cell Death, L-Type Calcium Channel, heart, Protein, PKB-ALPHA, Phenomena, Insulin A Chain, target of rapamycin complex, mHL-1, Sodium, Novolin, PKBBETA, metabolism, Isotope Coded Affinity Tagging, Mass Spectrum Analyses, flange, organ process, Metabolic Phenomenon, serine-threonine kinase Akt, apoptosis signaling, Mass Spectrum, Temperature, HIHGHH, mobilization, multicellular organism metabolic process, Calcium, hypertrophy, ASGP-R 1, apoptosis, biodegradation, Metabolic, Regulator, Kinases, INS, TOR signaling complex, Isotope Labeling, Hepatic lectin H1, Affinity Tagging, high voltage gated calcium channel activity, Isotopically-Coded, signalling, Phosphodiesterase, Collapse, process, processes, L Type Calcium Channels, L-Type VDCC beta Subunit, Herz@de, single-organism metabolic process, Coagulation Factor IV, Calcium-Dependent, L-Type Voltage-Dependent Calcium Channels, signalling process, apoptotic program, AW554154, PKBalpha, cardinality, Long-Lasting Calcium Channels, commitment to apoptosis, Isotope-Coded, processus, regulation, Phosphodiesterase Activator Protein, RAC, Rac, Receptor, Cyclic, Q-type calcium channel, quantitative, L Type VDCC gamma Subunit, RWDD5, L-Type VDCC, thymoma viral proto-oncogene, Anabolism, presence or absence in organism5705http://gpmdb.thegpm.org/~/dblist_gpmnum/gpmnum=GPM32310003337patentfamilies~0rfam~0merops~0complex-portal~0uniprot~0wormbaseparasite~0embl-covid19~0reactome~0emdb~0wgs_masters~0ebiweb_resources~0opentargets_genetics~0biomodels_all~0ipd-mhc~0ebiweb_teams~0taxonomy~0genome_assembly~0sc-experiments~0ebiweb_people~0enzymeportal_enzymes~0ipd-nhkir~0cellosaurus~0pdbe~0chebi~0patentproteins~0interpro7~0uniref~0chembl~0pdbekb~0gpcrdb~0hgnc~0sc-genes~0intact~0rhea~0ebiweb_training~0alphafold~0imgt-hla~0patentnucleotides~0ensemblroot~0eva_studies~0non-coding~0europepmc~0pubmed~1identifiers_registry~0pdbechem~0hpa-covid19~0eva-variants-covid19~0biosamples~0gwas_catalog~0biotools~0tls_masters~0mesh~0coding~0sra~0opentargets~0efo~0embl-pathogen~0project~0pride~1human_diseases~0geo_datasets~0embl~0treefam~0uniparc~0ols~0dgva~0intenz~0go~0tsa_masters~0biosamples-covid19~0ebiweb_corporate~0omim~0lrg~0earlycause-molecular-sequences~0ipd-kir~0empiar~0rnacentral~0orcid_data_claims~0gpmdb~2lineage-covid19~0metagenomics~0pfam~0pride archive~1varsite~00Resource Reanalysis0.00.00.01758173966687230.01.0falseAKT1 AND AKT2 INDUCE DISTINCT PHOSPHORYLATION PATTERNS IN HL-1 CARDIAC MYOCYTES.Data from ProteomeXchange, PXD ID: PXD000268. Experiment: DML2, file: folder summary. Published as part of J Proteome Res. 2014 Aug 27 . From the Abstract: {{i}} ... To gain more insight into AKT isoform specific signaling at the molecular level we investigated the phosphoproteome of HL-1 cardiomyocytes carrying AKT1 or AKT2 isoform specific knock down, respectively. We combined stable isotope labeling with high resolution mass spectrometry and identified 377 regulated phosphopeptides. Although AKT1 is expressed at four-fold higher levels, insulin stimulation mainly activated AKT2, which might in part rely on a preferred interaction of AKT2 with mammalian target of rapamycin complex 2. In line with this result, the highest number of regulated phosphopeptides was identified in the AKT2 knock down cells. ... {{/i}}2014-08-30GPM3231000333725162660PXD000268PXD000268