Mutagenesis of Thr-286 in monomeric Ca2+/calmodulin-dependent protein kinase II eliminates Ca2+/calmodulin-independent activity.
Open Access
- 1 February 1990
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 87 (4) , 1273-1277
- https://doi.org/10.1073/pnas.87.4.1273
Abstract
We have examined the role of Thr-286 autophosphorylation in the autoregulation of Ca2+/calmodulin-dependent protein kinase II. Using site-directed mutagenesis, we have substituted alanine or serine for Thr-286, or isoleucine for Arg-283, in the 50-kDa subunit of the kinase and expressed each protein in bacteria. Activation and autophosphorylation of all four enzymes were stringently dependent on Ca2+/calmodulin, indicating that neither Arg-283 nor Thr-286 is an absolute requirement for the pseudosubstrate inhibition of the enzyme. Autophosphorylation of the Ile-283 or Ala-286 enzyme generated little, if any, Ca2+/calmodulin-independent kinase activity, unlike the parent (Thr-286) or Ser-286 enzyme. The enzymes expressed in bacteria are predominantly monomeric, indicating that the generation of Ca2+/calmodulin-independent activity does not require the cooperative interactions of subunits normally present in the brain holoenzyme.Keywords
This publication has 30 references indexed in Scilit:
- Sequences of autophosphorylation sites in neuronal type II CaM kinase that control Ca2+-independent activityPublished by Elsevier ,1988
- Rabbit skeletal muscle myosin light chain kinase. The calmodulin binding domain as a potential active site-directed inhibitory domain.Journal of Biological Chemistry, 1987
- Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase.Proceedings of the National Academy of Sciences, 1987
- Ca2+/calmodulin-dependent protein kinase II: identification of autophosphorylation sites responsible for generation of Ca2+/calmodulin-independence.Proceedings of the National Academy of Sciences, 1987
- Functional Analysis of a Complementary DNA for the 50-Kilodalton Subunit of Calmodulin Kinase IIScience, 1987
- Deduced primary structure of the beta subunit of brain type II Ca2+/calmodulin-dependent protein kinase determined by molecular cloning.Proceedings of the National Academy of Sciences, 1987
- The calmodulin binding domain of chicken smooth muscle myosin light chain kinase contains a pseudosubstrate sequence.Journal of Biological Chemistry, 1987
- [52] Role of autophosphorylation in regulating calmodulin-dependent protein kinasesPublished by Elsevier ,1987
- Activation of the multifunctional Ca2+/calmodulin-dependent protein kinase by autophosphorylation: ATP modulates production of an autonomous enzyme.Proceedings of the National Academy of Sciences, 1986
- Autophosphorylation reversibly regulates the Ca2+/calmodulin-dependence of Ca2+/calmodulin-dependent protein kinase II.Proceedings of the National Academy of Sciences, 1986