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/ http://www.ebi.ac.uk/ chao-2011-1 A Mechanism for Tunable Autoinhibition in the Structure of a Human Ca(2+)/Calmodulin- Dependent Kinase II Holoenzyme. 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. kuriyan@berkeley.edu Cell (0092-8674) 2011 Only protein-protein interactions imex curation chao-2011-2 A Mechanism for Tunable Autoinhibition in the Structure of a Human Ca(2+)/Calmodulin- Dependent Kinase II Holoenzyme. 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. Autophosphorylation of CAMK2A was coupled to to the oxidation of NADPH, thus enabling a continual monitoring of the autophosphoarylation by spectrophotometry. kuriyan@berkeley.edu 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 calm_chick Calmodulin CALM CAM RCJMB04_24e7 protein protein chick Gallus gallus (Chicken) Chicken MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK calcium(2+) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) kcc2a-kcc2a 1182274 n/a 1182276 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 12.0 direct interaction direct interaction false false false Supp. table 1 3.5A Rworking=27.3% Rfree=32.7% calcium_2__-calm 1182275 n/a 1182277 cofactor cofactor coenzyme unspecified role unspecified role n/a 1182278 cofactor cofactor coenzyme unspecified role unspecified role ecoli Escherichia coli (strain K12) n/a 1182276 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 false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 1182275 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.