IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. Science signaling 2019 imex curation Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B., Menzel J., Kownatzki-Danger D., Tokar S., Ballone A., Unthan-Fechner K., Kilisch M., Lenz C., Urlaub H., Mori M., Ottmann C., Shattock MJ., Lehnart SE., Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de Accepted 2019-AUG-20 AT 09:50 BST AT 09:50 BST by PPORRAS EMBO Molecular Medicine 2019 Only protein-protein interactions imex curation imex curation added 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. Science signaling 2019 imex curation Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B., Menzel J., Kownatzki-Danger D., Tokar S., Ballone A., Unthan-Fechner K., Kilisch M., Lenz C., Urlaub H., Mori M., Ottmann C., Shattock MJ., Lehnart SE., Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de Accepted 2019-AUG-20 AT 09:50 BST AT 09:50 BST by PPORRAS EMBO Molecular Medicine 2019 Only protein-protein interactions imex curation imex curation added 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. Science signaling 2019 imex curation Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B., Menzel J., Kownatzki-Danger D., Tokar S., Ballone A., Unthan-Fechner K., Kilisch M., Lenz C., Urlaub H., Mori M., Ottmann C., Shattock MJ., Lehnart SE., Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de Accepted 2019-AUG-20 AT 09:50 BST AT 09:50 BST by PPORRAS EMBO Molecular Medicine 2019 Only protein-protein interactions imex curation imex curation added 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. 14-3-3 binding creates a memory of kinase action by stabilizing the modified state of phospholamban. Science signaling 2019 imex curation Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B., Menzel J., Kownatzki-Danger D., Tokar S., Ballone A., Unthan-Fechner K., Kilisch M., Lenz C., Urlaub H., Mori M., Ottmann C., Shattock MJ., Lehnart SE., Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation rat-cardiac_myocytes_neo rat cardiac myocytes neo neonatal_ventricular_myocyte neonatal ventricular myocyte proximity-dependent biotin identification proximity-dependent biotin identification sequence tag sequence tag identification sequence tag Menzel J., Kownatzki-Danger D., Tolkar S., Ballone A., Unthan-Fechner K., Kilisch M., Mori M., Ottmann C., Shattock MJ., Lehnart SE., and Schwappach B. 2019-07-29: Submitted by Blanche Schwappach, Institut fur Molekularbiologie Universitatsmedizin Gottingen, Gottingen, Germany submitted material can be found at smb://pcserv.ebi.ac.uk/sp-pro6/intact/local/data/curation-material/2019/Menzel The following info was provided by authors upon request: GST-PLN - sequence origin PLN: mus musculus (mouse), expressed in E.coli V5-APEX2-PLN - sequence origin APEX2: soybean-derived with the following mutations: K14D, W41F, E112K and A134P, sequence origin PLN: oryctolagus cuniculus , adenoviral transuction in neonatal rat cardiomyocytes Recombinant CAMKII - Isoform: delta, expressed in baculovirus (commercially available: Thermo Fisher Scientific, cat. PV3373) Recombinant PKA - catalytic subunit, expressed in E.coli (commercially available: New England Biolabs, cat. P6000) Recombinant 14-3-3 - sequence origin: human, expressed E.coli (same recombinant protein as MBP-14-3-3, the MBP tag was cleaved with Factor Xa enzyme) MBP-14-3-3 - sequence origin human, expressed E.coli Phosphatase assay experiment was not curated as no unique UniprotAC could be assigned to Alkaline phosphatase protein blanche.schwappach@med.uni-goettingen.de Accepted 2019-AUG-20 AT 09:50 BST AT 09:50 BST by PPORRAS EMBO Molecular Medicine 2019 Only protein-protein interactions imex curation imex curation added 1433e_rat 14-3-3 protein epsilon Ywhae Mitochondrial import stimulation factor L subunit protein protein rat Rattus norvegicus (Rat) Rat MDDREDLVYQAKLAEQAERYDEMVESMKKVAGMDVELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEENKGGEDKLKMIREYRQMVETELKLICCDILDVLDKHLIPAANTGESKVFYYKMKGDYHRYLAEFATGNDRKEAAENSLVAYKAASDIAMTELPPTHPIRLGLALNFSVFYYEILNSPDRACRLAKAAFDDAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDMQGDGEEQNKEALQDVEDENQ DIP protein P62260 original sequence version: 67 07817CCBD1F75B26 ppla_mouse Cardiac phospholamban Pln protein protein mouse Mus musculus Mouse MEKVQYLTRSAIRRASTIEMPQQARQNLQNLFINFCLILICLLLICIIVMLL 0763601F76A854D3 kcc2d_human Calcium/calmodulin-dependent protein kinase type II subunit delta CAMK2D CAMKD protein protein human Homo sapiens Human MASTTTCTRFTDEYQLFEELGKGAFSVVRRCMKIPTGQEYAAKIINTKKLSARDHQKLEREARICRLLKHPNIVRLHDSISEEGFHYLVFDLVTGGELFEDIVAREYYSEADASHCIQQILESVNHCHLNGIVHRDLKPENLLLASKSKGAAVKLADFGLAIEVQGDQQAWFGFAGTPGYLSPEVLRKDPYGKPVDMWACGVILYILLVGYPPFWDEDQHRLYQQIKAGAYDFPSPEWDTVTPEAKDLINKMLTINPAKRITASEALKHPWICQRSTVASMMHRQETVDCLKKFNARRKLKGAILTTMLATRNFSAAKSLLKKPDGVKESTESSNTTIEDEDVKARKQEIIKVTEQLIEAINNGDFEAYTKICDPGLTAFEPEALGNLVEGMDFHRFYFENALSKSNKPIHTIILNPHVHLVGDDAACIAYIRLTQYMDGSGMPKTMQSEETRVWHRRDGKWQNVHFHRSGSPTVPIKPPCIPNGKENFSGGTSLWQNI DIP protein Q13557 original sequence version: 123 DIP protein Q13557 original sequence version: 113 62999FBAB98120CE 1433f_rat 14-3-3 protein eta Ywhah protein protein rat Rattus norvegicus (Rat) Rat MGDREQLLQRARLAEQAERYDDMASAMKAVTELNEPLSNEDRNLLSVAYKNVVGARRSSWRVISSIEQKTMADGNEKKLEKVKAYREKIEKELETVCNDVLALLDKFLIKNCNDFQYESKVFYLKMKGDYYRYLAEVASGEKKNSVVEASEAAYKEAFEISKEHMQPTHPIRLGLALNFSVFYYEIQNAPEQACLLAKQAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDEEAGEGN DIP protein P68511 original sequence version: 56 C12F62B4ABA76DA3 1433t_rat 14-3-3 protein theta Ywhaq 14-3-3 protein tau protein protein rat Rattus norvegicus (Rat) Rat MEKTELIQKAKLAEQAERYDDMATCMKAVTEQGAELSNEERNLLSVAYKNVVGGRRSAWRVISSIEQKTDTSDKKLQLIKDYREKVESELRSICTTVLELLDKYLIANATNPESKVFYLKMKGDYFRYLAEVACGDDRKQTIENSQGAYQEAFDISKKEMQPTHPIRLGLALNFSVFYYEILNNPELACTLAKTAFDEAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDSAGEECDAAEGAEN E30471260E8B366E 1433g_rat 14-3-3 protein gamma Ywhag protein protein rat Rattus norvegicus (Rat) Rat MVDREQLVQKARLAEQAERYDDMAAAMKNVTELNEPLSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTSADGNEKKIEMVRAYREKIEKELEAVCQDVLSLLDNYLIKNCSETQYESKVFYLKMKGDYYRYLAEVATGEKRATVVESSEKAYSEAHEISKEHMQPTHPIRLGLALNYSVFYYEIQNAPEQACHLAKTAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDDDGGEGNN B0D16C6DE1F4455D ppla_rabit Cardiac phospholamban PLN protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MEKVQYLTRSAIRRASTIEMPQQARQNLQNLFINFCLILICLLLICIIVMLL DIP protein P61015 original sequence version: 37 DIP protein P61015 original sequence version: 67 DIP protein P61015 original sequence version: 73 0763601F76A854D3 1433g_human 14-3-3 protein gamma YWHAG Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MVDREQLVQKARLAEQAERYDDMAAAMKNVTELNEPLSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTSADGNEKKIEMVRAYREKIEKELEAVCQDVLSLLDNYLIKNCSETQYESKVFYLKMKGDYYRYLAEVATGEKRATVVESSEKAYSEAHEISKEHMQPTHPIRLGLALNYSVFYYEIQNAPEQACHLAKTAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDDDGGEGNN DIP protein P61981 original sequence version: 126 DIP protein P61981 original sequence version: 135 B0D16C6DE1F4455D 1433z_rat 14-3-3 protein zeta/delta Ywhaz Msfs1 Protein kinase C inhibitor protein 1 Mitochondrial import stimulation factor S1 subunit protein protein rat Rattus norvegicus (Rat) Rat MDKNELVQKAKLAEQAERYDDMAACMKSVTEQGAELSNEERNLLSVAYKNVVGARRSSWRVVSSIEQKTEGAEKKQQMAREYREKIETELRDICNDVLSLLEKFLIPNASQPESKVFYLKMKGDYYRYLAEVAAGDDKKGIVDQSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDTQGDEAEAGEGGEN DIP protein P63102 original sequence version: 65 2164DF3793B45B7A kapca_mouse cAMP-dependent protein kinase catalytic subunit alpha Prkaca Pkaca protein protein mouse Mus musculus Mouse MGNAAAAKKGSEQESVKEFLAKAKEDFLKKWETPSQNTAQLDQFDRIKTLGTGSFGRVMLVKHKESGNHYAMKILDKQKVVKLKQIEHTLNEKRILQAVNFPFLVKLEFSFKDNSNLYMVMEYVAGGEMFSHLRRIGRFSEPHARFYAAQIVLTFEYLHSLDLIYRDLKPENLLIDQQGYIQVTDFGFAKRVKGRTWTLCGTPEYLAPEIILSKGYNKAVDWWALGVLIYEMAAGYPPFFADQPIQIYEKIVSGKVRFPSHFSSDLKDLLRNLLQVDLTKRFGNLKNGVNDIKNHKWFATTDWIAIYQRKVEAPFIPKFKGPGDTSNFDDYEEEEIRVSINEKCGKEFTEF DIP protein P05132 original sequence version: 185 DIP protein P05132 original sequence version: 186 DIP protein P05132-1 original sequence version: 145 DIP protein P05132-1 original sequence version: 193 DIP protein P05132-1 original sequence version: 199 DIP protein P05132 original sequence version: 163 DIP protein P05132 original sequence version: 192 DIP protein P05132-1 original sequence version: 197 DIP protein P05132-1 original sequence version: 149 02F85D66EB21A1FA pln-1433 2 12 unspecified role unspecified role bait bait purified purified mbp tag mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey subcellular prep subcellular preparation subcellular prep cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level phosphorylated Ser opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain observed-ptm observed-ptm observed ptm observed-ptm phosphorylated Thr opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain observed-ptm observed-ptm observed ptm observed-ptm mouse-heart-ventricle Mouse heart ventricle Mouse_cardiac_ventricle Mouse Mouse heart ventricle Mouse cardiac ventricle heart_ventricle heart-ventricle physical association physical association Fig. 1E-I proteins specifically bound from the membrane fraction pln-10 4 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 13 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 5 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain APEX2 tag enzyme tag enzyme tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range v5 tag v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range proximity proximity close-contact colocalization neighbourhood interaction Fig. 2B, Data File S1 pln-1433g-1 2 14 unspecified role unspecified role ancillary ancillary purified purified ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P61014:p.Ser16Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 19:14:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser16Ala' P61014:p.Thr17Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain T A Tue Jun 30 15:40:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr17Ala' gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3C, 3D However, using the first 31 NT residues of PLN fused to GST we were unable to reconstitute 14-3-3 binding despite efficient phosphorylation of S16 by purified PKA (Fig 2A, S2). However, when we exchanged the neighboring residue T17 with an alanine, the affinity of pS16-PLN-GST for 14-3-3 was compatible with that of the binding assay (Fig 2A,C). This confirms that pS16 contributes to a 14-3-3 binding site that is, however, weakened by the neighboring T17. pln-ywhag-1 2 7 unspecified role unspecified role ancillary ancillary purified purified spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm 12 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P61014:p.Ser16Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 18:01:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser16Ala' P61014:p.Thr17Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain T A Tue Jun 30 19:04:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr17Ala' gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P61014:p.Arg9Cys mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R C Tue Jun 30 18:34:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg9Cys' P61014:p.Arg14del mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain R . Tue Jun 30 18:13:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg14del' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3G, 3H, 4D pln-ywhag-2 1 14 enzyme enzyme neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component purified purified P61014:p.Ser16Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 18:31:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser16Ala' P61014:p.Thr17Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain T A Tue Jun 30 18:17:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr17Ala' gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined phosphorylated Ser opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. S1C camk2d-pln-1 1 7 enzyme enzyme neutral component neutral component purified purified spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm 6 enzyme target enzyme target substrate substrate neutral component neutral component purified purified P61014:p.Ser16Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 16:40:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser16Ala' P61014:p.Thr17Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain T A Tue Jun 30 15:57:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr17Ala' gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined phosphorylated Thr opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. S1D pln-ywhag-3 3 14 unspecified role unspecified role ancillary ancillary purified purified ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait purified purified cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction Fig. S4 However, using the first 31 NT residues of PLN fused to GST we were unable to reconstitute 14-3-3 binding despite efficient phosphorylation of S16 by purified PKA (Fig 2A, S2). However, when we exchanged the neighboring residue T17 with an alanine, the affinity of pS16-PLN-GST for 14-3-3 was compatible with that of the binding assay (Fig 2A,C). This confirms that pS16 contributes to a 14-3-3 binding site that is, however, weakened by the neighboring T17.