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/ A Mechanism for Tunable Autoinhibition in the Structure of a Human Ca(2+)/Calmodulin- Dependent Kinase II Holoenzyme. A Mechanism for Tunable Autoinhibition in the Structure of a Human Ca(2+)/Calmodulin- Dependent Kinase II Holoenzyme. Cell (0092-8674) 2011 imex curation Chao L.H., Stratton M.M., Lee I.H., Rosenberg O.S., Levitz J., Mandell D.J., Kortemme T., Groves J.T., Schulman H., Kuriyan J. The description of the protein in the paper "We determined the crystal structure of an essentially full-length variant of the human β7 CaMKII holoenzyme" suggests the construct is CAMK2B. In fact this is due to an error, the phrase should read "..the crystallization construct α with a β7 linker (GenBank: NP_741960.1 and AAD42070.1, referred to as human β7)." The correct construct was deposited into PDB. Author assures me a correction will be printed by the journal. kuriyan@berkeley.edu 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Chao L.H., Stratton M.M., Lee I.H., Rosenberg O.S., Levitz J., Mandell D.J., Kortemme T., Groves J.T., Schulman H., Kuriyan J. The description of the protein in the paper "We determined the crystal structure of an essentially full-length variant of the human β7 CaMKII holoenzyme" suggests the construct is CAMK2B. In fact this is due to an error, the phrase should read "..the crystallization construct α with a β7 linker (GenBank: NP_741960.1 and AAD42070.1, referred to as human β7)." The correct construct was deposited into PDB. Author assures me a correction will be printed by the journal. kuriyan@berkeley.edu Accepted 2011-NOV-18 by JYOTI Cell (0092-8674) 2011 Only protein-protein interactions imex curation A Mechanism for Tunable Autoinhibition in the Structure of a Human Ca(2+)/Calmodulin- Dependent Kinase II Holoenzyme. A Mechanism for Tunable Autoinhibition in the Structure of a Human Ca(2+)/Calmodulin- Dependent Kinase II Holoenzyme. Cell (0092-8674) 2011 imex curation Chao L.H., Stratton M.M., Lee I.H., Rosenberg O.S., Levitz J., Mandell D.J., Kortemme T., Groves J.T., Schulman H., Kuriyan J. The description of the protein in the paper "We determined the crystal structure of an essentially full-length variant of the human β7 CaMKII holoenzyme" suggests the construct is CAMK2B. In fact this is due to an error, the phrase should read "..the crystallization construct α with a β7 linker (GenBank: NP_741960.1 and AAD42070.1, referred to as human β7)." The correct construct was deposited into PDB. Author assures me a correction will be printed by the journal. kuriyan@berkeley.edu 14-Mar-2012: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Chao L.H., Stratton M.M., Lee I.H., Rosenberg O.S., Levitz J., Mandell D.J., Kortemme T., Groves J.T., Schulman H., Kuriyan J. The description of the protein in the paper "We determined the crystal structure of an essentially full-length variant of the human β7 CaMKII holoenzyme" suggests the construct is CAMK2B. In fact this is due to an error, the phrase should read "..the crystallization construct α with a β7 linker (GenBank: NP_741960.1 and AAD42070.1, referred to as human β7)." The correct construct was deposited into PDB. Author assures me a correction will be printed by the journal. Autophosphorylation of CAMK2A was coupled to to the oxidation of NADPH, thus enabling a continual monitoring of the autophosphoarylation by spectrophotometry. kuriyan@berkeley.edu Accepted 2011-NOV-18 by JYOTI 14-Mar-2012: Contacted by JYOTI. Cell (0092-8674) 2011 Only protein-protein interactions imex curation kcc2a_human Calcium/calmodulin-dependent protein kinase type II subunit alpha CAMK2A CAMKA KIAA0968 protein protein human Homo sapiens Human MATITCTRFTEEYQLFEELGKGAFSVVRRCVKVLAGQEYAAKIINTKKLSARDHQKLEREARICRLLKHPNIVRLHDSISEEGHHYLIFDLVTGGELFEDIVAREYYSEADASHCIQQILEAVLHCHQMGVVHRDLKPENLLLASKLKGAAVKLADFGLAIEVEGEQQAWFGFAGTPGYLSPEVLRKDPYGKPVDLWACGVILYILLVGYPPFWDEDQHRLYQQIKAGAYDFPSPEWDTVTPEAKDLINKMLTINPSKRITAAEALKHPWISHRSTVASCMHRQETVDCLKKFNARRKLKGAILTTMLATRNFSGGKSGGNKKSDGVKESSESTNTTIEDEDTKVRKQEIIKVTEQLIEAISNGDFESYTKMCDPGMTAFEPEALGNLVEGLDFHRFYFENLWSRNSKPVHTTILNPHIHLMGDESACIAYIRITQYLDAGGIPRTAQSEETRVWHRRDGKWQIVHFHRSGAPSVLPH DIP protein Q9UQM7 original sequence version: 96 208143A311BA9262 calm_chick Calmodulin CALM CAM RCJMB04_24e7 protein protein chick Gallus gallus (Chicken) Chicken MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK DIP protein P62149 original sequence version: 32 DIP protein P62149 original sequence version: 64 6B4BC3FCDE10727B calcium(2+) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) kcc2a-kcc2a 1 3 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp. table 1 3.5A Rworking=27.3% Rfree=32.7% calcium_2__-calm 2 4 cofactor cofactor coenzyme unspecified role unspecified role 5 cofactor cofactor coenzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) 3 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 4A, 4B Supp Fig. 4 The human short-linker CaMKII shows high cooperativity and a microM EC50 value, whereas a human short-linker CaMKII with central hub mutation (ile351glu) shows non-cooperative activation and a nanoM EC50 value. Increasing the linker length increases the calcium-independent activity of the molecule. Mutations thr370glu and leu392glu abrogates cooperative activation and shifts the EC50 to lower calmodulin concentrations. The Hill coefficient increases when the concentration of protein in the assay increases to mimic the 'crowding' conditions of a cell.