Cytoskeletal-like Filaments of Ca2+-Calmodulin-Dependent Protein Kinase II Are Formed in a Regulated and Zn2+-Dependent Manner.
Ca2+-calmodulin-dependent protein kinase II (CaMKII) is highly abundant in neurons, where its concentration reaches that typically found for cytoskeletal proteins. Functional reasons for such a high concentration are not known, but given the multitude of known binding partners for CaMKII, a role as a scaffolding molecule has been proposed. In this report, we provide experimental evidence that demonstrates a novel structural role for CaMKII. We discovered that CaMKII forms filaments that can extend for several micrometers in the presence of certain divalent cations (Zn2+, Cd2+, and Cu2+) but not with others (Ca2+, Mg2+, Co2+, and Ni2+). Once formed, depleting the divalent ion concentration with chelators completely dissociated the filaments, and this process could be repeated by cyclic addition and removal of divalent ions. Using the crystal structure of the CaMKII holoenzyme, we computed an electrostatic potential map of the dodecameric complex to predict divalent ion binding sites. This analysis revealed a potential surface-exposed divalent ion binding site involving amino acids that also participate in calmodulin (CaM) binding and suggested CaM binding might inhibit formation of the filaments. As predicted, Ca2+/CaM binding both inhibited divalent ion-induced filament formation and could disassemble preformed filaments. Interestingly, CaMKII within the filaments retains the capacity to autophosphorylate; however, activity toward exogenous substrates is significantly decreased. Activity is restored upon filament disassembly. We compile our results with structural and mechanistic data from the literature to propose a model of Zn2+-mediated CaMKII filament formation, in which assembly and activity are further regulated by Ca2+/CaM.
Department of Neurobiology and Anatomy, McGovern Medical School, The University of Texas Health Science Center at Houston , 6431 Fannin, Room 7.254, MSB, Houston, Texas 77030, United States.
Laurel
L
Hoffman
Department of Physics and Astronomy, Clemson University , Clemson, South Carolina 29634-0978, United States.
Lin
L
Li
Department of Physics and Astronomy, Clemson University , Clemson, South Carolina 29634-0978, United States.
Emil
0000-0001-5346-0156
E
Alexov
Department of Physics and Astronomy, Clemson University , Clemson, South Carolina 29634-0978, United States.
Hugo
0000-0001-7068-6827
H
Sanabria
Department of Neurobiology and Anatomy, McGovern Medical School, The University of Texas Health Science Center at Houston , 6431 Fannin, Room 7.254, MSB, Houston, Texas 77030, United States.
M Neal
0000-0003-4801-1190
MN
Waxham
D000818:Animals; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000737:chemistry*; D002460:Cell Line; D003599:Cytoskeleton / Q000737:chemistry*; D007700:Kinetics; D010766:Phosphorylation; D051381:Rats; D018411:Spodoptera; D055672:Static Electricity; D015032:Zinc / Q000737:chemistry*
D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D015032:Zinc