PMID- 32902386 OWN - NLM STAT- MEDLINE DCOM- 20210223 LR - 20210223 IS - 2050-084X (Electronic) IS - 2050-084X (Linking) VI - 9 DP - 2020 Sep 9 TI - Breakage of the oligomeric CaMKII hub by the regulatory segment of the kinase. LID - 10.7554/eLife.57784 [doi] LID - e57784 AB - Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is an oligomeric enzyme with crucial roles in neuronal signaling and cardiac function. Previously, we showed that activation of CaMKII triggers the exchange of subunits between holoenzymes, potentially increasing the spread of the active state (Stratton et al., 2014; Bhattacharyya et al., 2016). Using mass spectrometry, we show now that unphosphorylated and phosphorylated peptides derived from the CaMKII-alpha regulatory segment bind to the CaMKII-alpha hub and break it into smaller oligomers. Molecular dynamics simulations show that the regulatory segments dock spontaneously at the interface between hub subunits, trapping large fluctuations in hub structure. Single-molecule fluorescence intensity analysis of CaMKII-alpha expressed in mammalian cells shows that activation of CaMKII-alpha results in the destabilization of the holoenzyme. Our results suggest that release of the regulatory segment by activation and phosphorylation allows it to destabilize the hub, producing smaller assemblies that might reassemble to form new holoenzymes. CI - (c) 2020, Karandur et al. FAU - Karandur, Deepti AU - Karandur D AUID- ORCID: 0000-0002-6949-6337 AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States. FAU - Bhattacharyya, Moitrayee AU - Bhattacharyya M AUID- ORCID: 0000-0002-2168-1541 AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States. FAU - Xia, Zijie AU - Xia Z AD - Department of Chemistry, University of California, Berkeley, Berkeley, United States. FAU - Lee, Young Kwang AU - Lee YK AUID- ORCID: 0000-0003-0056-6357 AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Department of Chemistry, University of California, Berkeley, Berkeley, United States. FAU - Muratcioglu, Serena AU - Muratcioglu S AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States. FAU - McAffee, Darren AU - McAffee D AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Department of Chemistry, University of California, Berkeley, Berkeley, United States. FAU - McSpadden, Ethan D AU - McSpadden ED AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States. FAU - Qiu, Baiyu AU - Qiu B AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States. FAU - Groves, Jay T AU - Groves JT AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Department of Chemistry, University of California, Berkeley, Berkeley, United States. AD - Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, United States. FAU - Williams, Evan R AU - Williams ER AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Department of Chemistry, University of California, Berkeley, Berkeley, United States. FAU - Kuriyan, John AU - Kuriyan J AUID- ORCID: 0000-0002-4414-5477 AD - Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, United States. AD - Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States. AD - Department of Chemistry, University of California, Berkeley, Berkeley, United States. AD - Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, United States. LA - eng GR - R00 GM126145/GM/NIGMS NIH HHS/United States GR - CHE-1609866/National Science Foundation/International GR - HHMI/Howard Hughes Medical Institute/United States GR - R01 GM116961/GM/NIGMS NIH HHS/United States GR - K99 GM126145/GM/NIGMS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20200909 PL - England TA - Elife JT - eLife JID - 101579614 RN - 0 (CAMK2N1 protein, human) RN - 0 (Holoenzymes) RN - 0 (Proteins) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 2) SB - IM MH - Calcium-Calmodulin-Dependent Protein Kinase Type 2/*antagonists & inhibitors MH - Escherichia coli MH - HEK293 Cells MH - Holoenzymes/metabolism MH - Humans MH - Molecular Dynamics Simulation MH - Phosphorylation MH - Proteins/*genetics/metabolism MH - Signal Transduction/genetics PMC - PMC7538161 OTO - NOTNLM OT - CaMKII OT - activation-dependent disassembly OT - biochemistry OT - chemical biology OT - molecular dynamics simulation OT - native mass spectrometry OT - spread of activation state COIS- DK, MB, ZX, YL, SM, DM, EM, BQ, JG, EW No competing interests declared, JK Senior editor, eLife EDAT- 2020/09/10 06:00 MHDA- 2021/02/24 06:00 PMCR- 2020/09/09 CRDT- 2020/09/09 12:23 PHST- 2020/04/15 00:00 [received] PHST- 2020/09/08 00:00 [accepted] PHST- 2020/09/10 06:00 [pubmed] PHST- 2021/02/24 06:00 [medline] PHST- 2020/09/09 12:23 [entrez] PHST- 2020/09/09 00:00 [pmc-release] AID - 57784 [pii] AID - 10.7554/eLife.57784 [doi] PST - epublish SO - Elife. 2020 Sep 9;9:e57784. doi: 10.7554/eLife.57784.