PMID- 30942928 OWN - NLM STAT- MEDLINE DCOM- 20200410 LR - 20200601 IS - 1469-896X (Electronic) IS - 0961-8368 (Print) IS - 0961-8368 (Linking) VI - 28 IP - 6 DP - 2019 Jun TI - Variation in assembly stoichiometry in non-metazoan homologs of the hub domain of Ca(2+) /calmodulin-dependent protein kinase II. PG - 1071-1082 LID - 10.1002/pro.3614 [doi] AB - The multi-subunit Ca(2+) /calmodulin-dependent protein kinase II (CaMKII) holoenzyme plays a critical role in animal learning and memory. The kinase domain of CaMKII is connected by a flexible linker to a C-terminal hub domain that assembles into a 12- or 14-subunit scaffold that displays the kinase domains around it. Studies on CaMKII suggest that the stoichiometry and dynamic assembly/disassembly of hub oligomers may be important for CaMKII regulation. Although CaMKII is a metazoan protein, genes encoding predicted CaMKII-like hub domains, without associated kinase domains, are found in the genomes of some green plants and bacteria. We show that the hub domains encoded by three related green algae, Chlamydomonas reinhardtii, Volvox carteri f. nagarensis, and Gonium pectoral, assemble into 16-, 18-, and 20-subunit oligomers, as assayed by native protein mass spectrometry. These are the largest known CaMKII hub domain assemblies. A crystal structure of the hub domain from C. reinhardtii reveals an 18-subunit organization. We identified four intra-subunit hydrogen bonds in the core of the fold that are present in the Chlamydomonas hub domain, but not in metazoan hubs. When six point mutations designed to recapitulate these hydrogen bonds were introduced into the human CaMKII-alpha hub domain, the mutant protein formed assemblies with 14 and 16 subunits, instead of the normal 12- and 14-subunit assemblies. Our results show that the stoichiometric balance of CaMKII hub assemblies can be shifted readily by small changes in sequence. CI - (c) 2019 The Protein Society. FAU - McSpadden, Ethan D AU - McSpadden ED AD - Department of Molecular and Cell Biology, University of California, Berkeley, California. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California. AD - Howard Hughes Medical Institute, University of California, Berkeley, California. FAU - Xia, Zijie AU - Xia Z AD - Department of Chemistry, University of California, Berkeley, California. FAU - Chi, Chris C AU - Chi CC AD - Department of Molecular and Cell Biology, University of California, Berkeley, California. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California. AD - Howard Hughes Medical Institute, University of California, Berkeley, California. FAU - Susa, Anna C AU - Susa AC AD - Department of Chemistry, University of California, Berkeley, California. FAU - Shah, Neel H AU - Shah NH AUID- ORCID: 0000-0002-1186-0626 AD - Department of Molecular and Cell Biology, University of California, Berkeley, California. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California. AD - Howard Hughes Medical Institute, University of California, Berkeley, California. FAU - Gee, Christine L AU - Gee CL AUID- ORCID: 0000-0002-2632-6418 AD - Department of Molecular and Cell Biology, University of California, Berkeley, California. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California. AD - Howard Hughes Medical Institute, University of California, Berkeley, California. FAU - Williams, Evan R AU - Williams ER AUID- ORCID: 0000-0002-1733-3018 AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California. AD - Department of Chemistry, University of California, Berkeley, California. FAU - Kuriyan, John AU - Kuriyan J AUID- ORCID: 0000-0002-4414-5477 AD - Department of Molecular and Cell Biology, University of California, Berkeley, California. AD - California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, California. AD - Howard Hughes Medical Institute, University of California, Berkeley, California. AD - Department of Chemistry, University of California, Berkeley, California. AD - Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California. LA - eng GR - HHMI/Howard Hughes Medical Institute/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20190417 PL - United States TA - Protein Sci JT - Protein science : a publication of the Protein Society JID - 9211750 RN - 0 (Protein Subunits) RN - EC 2.7.11.17 (Calcium-Calmodulin-Dependent Protein Kinase Type 2) SB - IM MH - Amino Acid Sequence MH - Calcium-Calmodulin-Dependent Protein Kinase Type 2/*chemistry/isolation & purification/metabolism MH - Crystallography, X-Ray MH - Humans MH - Models, Molecular MH - Protein Domains MH - Protein Subunits/chemistry/isolation & purification/metabolism MH - Sequence Alignment PMC - PMC6511747 OTO - NOTNLM OT - Chlamydomonas reinhardtii OT - CaMKII OT - X-ray crystallography OT - evolution OT - native protein mass spectrometry OT - oligomerization OT - protein engineering COIS- The authors declare that they have no conflict of interest. EDAT- 2019/04/04 06:00 MHDA- 2020/04/11 06:00 PMCR- 2020/06/01 CRDT- 2019/04/04 06:00 PHST- 2019/01/30 00:00 [received] PHST- 2019/04/01 00:00 [accepted] PHST- 2019/04/04 06:00 [pubmed] PHST- 2020/04/11 06:00 [medline] PHST- 2019/04/04 06:00 [entrez] PHST- 2020/06/01 00:00 [pmc-release] AID - PRO3614 [pii] AID - 10.1002/pro.3614 [doi] PST - ppublish SO - Protein Sci. 2019 Jun;28(6):1071-1082. doi: 10.1002/pro.3614. Epub 2019 Apr 17.