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PMID: 1334558 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of intrasteric inhibition of the multifunctional calcium/calmodulin-dependent protein kinase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 24 ·1992-12-15 ·Pages 12127-31

Cruzalegui FH, Kapiloff MS, Morfin JP, Kemp BE, Rosenfeld MG, Means AR

Abstract

A regulatory region involved in both autoinhibition and calmodulin (CaM) binding has previously been identified in the multifunctional Ca2+/CaM-dependent protein kinase (CaM kinase II). We have tested the role of various segments of the regulatory region in autoinhibition by the analysis of a series of truncation, substitution, and deletion mutants of the CaM kinase II alpha subunit (CaM kinase II alpha). Unexpectedly, the sequence Lys-Lys-Phe-Asn at positions 291-294, adjacent to the CaM binding domain, was found to be sufficient to maintain an inhibited state in a truncated form of the kinase. However, these residues are not essential in the context of the full-length protein, indicating the importance of additional residues from the overlapping CaM binding domain. We propose here a molecular model for CaM kinase II alpha based on the three-dimensional structure of the cAPK-PKI-(5-24) (protein kinase inhibitor fragment) complex. It is predicted from this model that autoinhibition is of the pseudosubstrate variety and that autophosphorylation of Thr-286 could occur by an intersubunit reaction in the holoenzyme complex.

MeSH Terms
Allosteric Regulation Amino Acid Sequence Calcium-Calmodulin-Dependent Protein Kinases Calmodulin-Binding Proteins/chemistry,metabolism,ultrastructure Computer Simulation DNA Mutational Analysis Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Kinase Inhibitors Protein Kinases/chemistry,metabolism Protein Structure, Tertiary Sequence Alignment Sequence Deletion Structure-Activity Relationship
Chemicals
Calmodulin-Binding Proteins Protein Kinase Inhibitors Protein Kinases Calcium-Calmodulin-Dependent Protein Kinases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Cruzalegui F H
Department of Pharmacology, Duke University Medical Center, Durham, NC 27710.
Kapiloff M S
Morfin J P
Kemp B E
Rosenfeld M G
Means A R
References (38)
38 references, click to expand
  1. Calmodulin trapping by calcium-calmodulin-dependent protein kinase.
    Science. 1992 May 22;256(5060):1199-202 PMID: 1317063
  2. Substrate specificity of a multifunctional calmodulin-dependent protein kinase.
    J Biol Chem. 1985 Nov 25;260(27):14471-6 PMID: 4055784
  3. Expression of a constitutive form of calcium/calmodulin dependent protein kinase II leads to arrest of the cell cycle in G2.
    EMBO J. 1992 Feb;11(2):507-17 PMID: 1371461
  4. Solution structure of a calmodulin-target peptide complex by multidimensional NMR.
    Science. 1992 May 1;256(5057):632-8 PMID: 1585175
  5. Active catalytic fragment of Ca2+/calmodulin-dependent protein kinase II. Purification, characterization, and structural analysis.
    J Biol Chem. 1991 Aug 15;266(23):15391-7 PMID: 1651329
  6. Identification of amino acids essential for calmodulin binding and activation of smooth muscle myosin light chain kinase.
    J Biol Chem. 1992 Feb 15;267(5):3024-9 PMID: 1737757
  7. Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase.
    Science. 1991 Jul 26;253(5018):414-20 PMID: 1862343
  8. Mutagenesis of Thr-286 in monomeric Ca2+/calmodulin-dependent protein kinase II eliminates Ca2+/calmodulin-independent activity.
    Proc Natl Acad Sci U S A. 1990 Feb;87(4):1273-7 PMID: 2154738
  9. Use of T7 RNA polymerase to direct expression of cloned genes.
    Methods Enzymol. 1990;185:60-89 PMID: 2199796
  10. Regulatory domain of calcium/calmodulin-dependent protein kinase II. Mechanism of inhibition and regulation by phosphorylation.
    J Biol Chem. 1989 Mar 25;264(9):4800-4 PMID: 2538462
  11. Expression and characterization of calmodulin-dependent protein kinase II from cloned cDNAs in Chinese hamster ovary cells.
    J Biol Chem. 1989 Nov 15;264(32):19108-16 PMID: 2553731
  12. Primary structural determinants essential for potent inhibition of cAMP-dependent protein kinase by inhibitory peptides corresponding to the active portion of the heat-stable inhibitor protein.
    J Biol Chem. 1989 May 25;264(15):8802-10 PMID: 2722799
  13. Regulatory interactions of the calmodulin-binding, inhibitory, and autophosphorylation domains of Ca2+/calmodulin-dependent protein kinase II.
    J Biol Chem. 1988 Dec 5;263(34):18145-51 PMID: 2848027
  14. Functional analysis of a complementary DNA for the 50-kilodalton subunit of calmodulin kinase II.
    Science. 1987 Jul 17;237(4812):293-7 PMID: 3037704
  15. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074-8 PMID: 3156376
  16. Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase.
    Proc Natl Acad Sci U S A. 1987 Aug;84(16):5962-6 PMID: 3475713
  17. Protein kinase C contains a pseudosubstrate prototope in its regulatory domain.
    Science. 1987 Dec 18;238(4834):1726-8 PMID: 3686012
  18. Structural basis of the intrasteric regulation of myosin light chain kinases.
    Science. 1992 Oct 2;258(5079):130-5 PMID: 1439761
  19. Intrasteric regulation of myosin light chain kinase: the pseudosubstrate prototope binds to the active site.
    Mol Endocrinol. 1992 Apr;6(4):621-6 PMID: 1584224
  20. A novel Ca2+/calmodulin-dependent protein kinase and a male germ cell-specific calmodulin-binding protein are derived from the same gene.
    Mol Cell Biol. 1991 Aug;11(8):3960-71 PMID: 1649385
  21. Intrasteric regulation of protein kinases and phosphatases.
    Biochim Biophys Acta. 1991 Aug 13;1094(1):67-76 PMID: 1653024
  22. Studies on the regulatory domain of Ca2+/calmodulin-dependent protein kinase II by expression of mutated cDNAs in Escherichia coli.
    J Biol Chem. 1991 Sep 5;266(25):16401-8 PMID: 1653235
  23. Crystal structure of the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase.
    Science. 1991 Jul 26;253(5018):407-14 PMID: 1862342
  24. Studies on the regulatory domain of Ca2+/calmodulin-dependent protein kinase II. Functional analyses of arginine 283 using synthetic inhibitory peptides and site-directed mutagenesis of the alpha subunit.
    J Biol Chem. 1990 Jul 5;265(19):11091-7 PMID: 1972705
  25. Multifunctional Ca2+/calmodulin-dependent protein kinase made Ca2+ independent for functional studies.
    Biochemistry. 1990 Feb 20;29(7):1679-84 PMID: 2158811
  26. Expression and characterization of the alpha-subunit of Ca2+/calmodulin-dependent protein kinase II using the baculovirus expression system.
    Biochem Biophys Res Commun. 1990 Dec 14;173(2):578-84 PMID: 2175600
  27. Functional analysis of Ca2+/calmodulin-dependent protein kinase II expressed in bacteria.
    J Biol Chem. 1989 May 5;264(13):7477-82 PMID: 2496129
  28. Multifunctional Ca2+/calmodulin-dependent protein kinase: domain structure and regulation.
    Trends Biochem Sci. 1989 Feb;14(2):62-6 PMID: 2539662
  29. Studies of the regulatory mechanism of Ca2+/calmodulin-dependent protein kinase II. Mutation of threonine 286 to alanine and aspartate.
    J Biol Chem. 1989 Oct 5;264(28):16759-63 PMID: 2550458
  30. cAMP-dependent protein kinase. Model for an enzyme family.
    J Biol Chem. 1989 May 25;264(15):8443-6 PMID: 2656679
  31. Calcium/calmodulin-dependent protein kinase II. Characterization of distinct calmodulin binding and inhibitory domains.
    J Biol Chem. 1988 May 25;263(15):7190-5 PMID: 2835367
  32. The multifunctional Ca2+/calmodulin-dependent protein kinase.
    Adv Second Messenger Phosphoprotein Res. 1988;22:39-112 PMID: 2852025
  33. Sequences of autophosphorylation sites in neuronal type II CaM kinase that control Ca2(+)-independent activity.
    Neuron. 1988 Sep;1(7):593-604 PMID: 2856100
  34. Active site-directed inhibition of Ca2+/calmodulin-dependent protein kinase type II by a bifunctional calmodulin-binding peptide.
    Proc Natl Acad Sci U S A. 1988 Jul;85(14):4991-5 PMID: 3134656
  35. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains.
    Science. 1988 Jul 1;241(4861):42-52 PMID: 3291115
  36. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Methods Enzymol. 1987;154:367-82 PMID: 3323813
  37. Rabbit skeletal muscle myosin light chain kinase. The calmodulin binding domain as a potential active site-directed inhibitory domain.
    J Biol Chem. 1987 Sep 5;262(25):11958-63 PMID: 3624242
  38. Inhibitory autophosphorylation of multifunctional Ca2+/calmodulin-dependent protein kinase analyzed by site-directed mutagenesis.
    J Biol Chem. 1992 Aug 25;267(24):17216-24 PMID: 1324926
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.24.12127
Published
1992-12-15
Pages
12127-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50711
Subset
IM
Grants
NIGMS NIH HHS · GM33976 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.24.12127 PMC Full Text View journal details
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