Cited by 94Protein Kinase Regulation and GTPase SignalingProtein Structure and DynamicsS100 Proteins and Annexins

A Peptide Model for Calmodulin Trapping by Calcium/Calmodulin-dependent Protein Kinase II

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Putkey, John A., Waxham, M. Neal·1996·Journal of Biological Chemistry

Abstract

Autophosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaM-kinase) induces a more than 1000-fold increase in calmodulin (CaM)-binding affinity by dramatically decreasing the off-rate for CaM. In this report, we investigate the molecular mechanism for this phenomenon by comparing the rate of dissociation of a novel fluorescently labeled CaM from two synthetic peptides and from the phosphorylated and nonphosphorylated forms of a recombinant preparation of CaM-kinase. Dissociation of a complex of CaM and CKII(296-312), a peptide representing close to the minimum CaM-binding domain of the alpha subunit of CaM-kinase, exhibited a fast off-rate of 5.0 s-1. This was similar to the off-rate of 1.1 s-1 for the dissociation of CaM from the nonphosphorylated form of CaM-kinase. In contrast, dissociation of CaM from either autophosphorylated CaM-kinase or peptide CKII(290-314) was extremely slow with apparent off-rates of about 3-9 x 10(-5) s-1. Along with information from the crystal structure of Ca2+/CaM bound to CKII(290-314) (Meador, W. E., Means, A. R., and Quiocho, F. A. (1993) Science 262, 1718-1721), our results suggest a model in which CaM-dependent autophosphorylation of CaM-kinase induces a conformational change in the region of the CaM-binding domain which allows the formation of additional stabilizing interactions with CaM. We predict that this involves amino acids 293-298 in CaM-kinase. The possible consequences of these observations on the reversibility of CaM…

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Crossref · 10.1074/jbc.271.47.29619

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  • The increased affinity for Ca2+/CaM is due to a greatly reduced dissociation rate of Ca2+/CaM, referred to as CaM trapping.

    “The increased affinity is due to a greatly reduced dissociation rate of Ca2+/CaM; it takes many seconds to dissociate when Ca2+ is elevated versus a fraction of a second before autophosphorylation and is referred to as CaM trapping (91, 92).”

    From Synaptic memory and CaMKII (2023) · cites this paper as [92]

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Authors
Putkey, J. A., & Waxham, M. N.
Corresponding author
Putkey, John A.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1996-11
Volume
271
Issue
47
Pages
29619-29623
ISSN
0021-9258 (ISSN-L)
OpenAlex
W1977861693
Cited by
94
References
31
Citations by year
2026: 2 · 2025: 5 · 2024: 3 · 2023: 5 · 2020: 5 · 2019: 3 · 2018: 3 · 2017: 2 · 2016: 2 · 2015: 1 · 2014: 7 · 2013: 3 · 2012: 1
FWCI
1.15
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (5r01hl045724-09, 5k04ns001509-03)
  • National Heart, Lung, and Blood Institute (HL45724)
  • National Institute of Neurological Disorders and Stroke (NS-26086, NS01509)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › S100 Proteins and Annexins
Keywords
calcium/calmodulin-dependent protein kinase II, calmodulin-binding domain, model peptides, conformational change
MeSH terms
Animals, Calmodulin, Kinetics, Models, Chemical, Peptides, Phosphorylation, Protein Binding, Recombinant Proteins, Calcium-Calmodulin-Dependent Protein Kinases, Rats, Calcium-Calmodulin-Dependent Protein Kinase Type 2

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