» Articles » PMID: 2162839

Calcium/calmodulin-independent Autophosphorylation Sites of Calcium/calmodulin-dependent Protein Kinase II. Studies on the Effect of Phosphorylation of Threonine 305/306 and Serine 314 on Calmodulin Binding Using Synthetic Peptides

Overview
Journal J Biol Chem
Specialty Biochemistry
Date 1990 Jul 5
PMID 2162839
Citations 55
Authors
Affiliations
Soon will be listed here.
Abstract

Two synthetic peptides containing the previously identified calmodulin (CaM)-binding domain of Ca2+/CaM-dependent protein kinase II (CaM-kinase II) (residues 296-309, Payne, M. E., Fong, Y.-L., Ono, T., Colbran, R. J., Kemp, B. E., Soderling, T. R., and Means, A. R. (1988) J. Biol. Chem. 263, 7190-7195) were phosphorylated by Ca2+/CaM-independent forms of the kinase. In the presence of EGTA, CaMK-(290-309) was phosphorylated exclusively on threonine residues (Km = 13 microM; Vmax = 211 nmol/min/mg). When the phosphorylated product was analyzed by reversed-phase high performance liquid chromatography (HPLC) two radioactive peaks were resolved. The first peak contained CaMK-(290-309) phosphorylated on Thr306, whereas the second peak contained CaMK-(290-309) phosphorylated on Thr305. However, under the same conditions CaMK-(294-319) was phosphorylated predominantly (approximately 70%) on serine residues (Km = 23 microM; Vmax = 99 nmol/min/mg) and HPLC analysis revealed a single major radioactive peak predominantly (more than 90%) phosphorylated at Ser314. Phosphorylation of both peptides was completely blocked in the presence of Ca2+ and a stoichiometric amount of CaM. Samples of each phosphorylated peptide were tested for CaM-binding ability by two procedures and compared to the nonphosphorylated peptides. Phosphorylation of either Thr305 or Thr306 greatly reduced the interaction between CaMK-(290-309) and CaM, whereas phosphorylation of Ser314 did not affect the ability of CaMK-(294-319) to bind CaM. These results indicate that Thr305 and/or Thr306 may be the Ca2+/CaM-independent autophosphorylation site(s) responsible for the loss of ability of CaM-kinase II to bind and be activated by Ca2+/CaM (Hashimoto, Y., Schworer, C. M., Colbran, R. J., and Soderling, T. R., J. Biol. Chem. 262, 8051-8055).

Citing Articles

Imaging single CaMKII holoenzymes at work by high-speed atomic force microscopy.

Tsujioka S, Sumino A, Nagasawa Y, Sumikama T, Flechsig H, Puppulin L Sci Adv. 2023; 9(26):eadh1069.

PMID: 37390213 PMC: 10313165. DOI: 10.1126/sciadv.adh1069.


Calcium/Calmodulin-Stimulated Protein Kinase II (CaMKII): Different Functional Outcomes from Activation, Depending on the Cellular Microenvironment.

Rostas J, Skelding K Cells. 2023; 12(3).

PMID: 36766743 PMC: 9913510. DOI: 10.3390/cells12030401.


The role of calcium and CaMKII in sleep.

Wang Y, Minami Y, Ode K, Ueda H Front Syst Neurosci. 2023; 16:1059421.

PMID: 36618010 PMC: 9815122. DOI: 10.3389/fnsys.2022.1059421.


Distinct phosphorylation states of mammalian CaMKIIβ control the induction and maintenance of sleep.

Tone D, Ode K, Zhang Q, Fujishima H, Yamada R, Nagashima Y PLoS Biol. 2022; 20(10):e3001813.

PMID: 36194579 PMC: 9531794. DOI: 10.1371/journal.pbio.3001813.


CaMKII as a Therapeutic Target in Cardiovascular Disease.

Reyes Gaido O, Nkashama L, Schole K, Wang Q, Umapathi P, Mesubi O Annu Rev Pharmacol Toxicol. 2022; 63:249-272.

PMID: 35973713 PMC: 11019858. DOI: 10.1146/annurev-pharmtox-051421-111814.