Biomaterial Database

Purification and characterization of Ca2+/calmodulin-dependent protein kinase V from rat cerebrum. 1993

H Mochizuki, and T Ito, and H Hidaka

Department of Pharmacology, Nagoya University School of Medicine, Japan.

A novel Ca2+/calmodulin-dependent protein kinase (CaM kinase V) from rat cerebrum was purified. This kinase phosphorylates the synthetic peptide substrate syntide-2. The purified enzyme showed a single protein band with a molecular mass of 41 kDa on SDS-polyacrylamide gel electrophoresis. The Stokes radius and the sedimentation coefficient were 31.8 A and 2.83 S, respectively. An approximate molecular mass of 37 kDa was calculated for the native enzyme, and a monomeric structure of the enzyme was suggested. Expression of the enzymatic activity required the presence of both Ca2+ and calmodulin (apparent Ka = 24 +/- 7 nM). The CaM kinase V had an apparent Km for ATP of 75 +/- 11 microM and for syntide-2 of 20 +/- 4 microM. CaM kinase V undergoes autophosphorylation in response to Ca2+ and calmodulin. CaM kinase V was digested with lysyl endopeptidase, and the partial amino acid sequence was determined. A computer homology search revealed no identical protein. KN-62, a selective inhibitor of Ca2+/calmodulin-dependent protein kinase II, inhibited CaM kinase V, with a Ki of 0.8 microM. CaM kinase V phosphorylates a number of endogenous proteins.

UI	MeSH Term	Description	Entries
D007546	Isoquinolines	A group of compounds with the heterocyclic ring structure of benzo(c)pyridine. The ring structure is characteristic of the group of opium alkaloids such as papaverine. (From Stedman, 25th ed)
D008297	Male		Males
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D010455	Peptides	Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are considered to be larger versions of peptides that can form into complex structures such as ENZYMES and RECEPTORS.	Peptide,Polypeptide,Polypeptides
D010766	Phosphorylation	The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.	Phosphorylations
D010879	Piperazines	Compounds that are derived from PIPERAZINE.
D001921	Brain	The part of CENTRAL NERVOUS SYSTEM that is contained within the skull (CRANIUM). Arising from the NEURAL TUBE, the embryonic brain is comprised of three major parts including PROSENCEPHALON (the forebrain); MESENCEPHALON (the midbrain); and RHOMBENCEPHALON (the hindbrain). The developed brain consists of CEREBRUM; CEREBELLUM; and other structures in the BRAIN STEM.	Encephalon
D002850	Chromatography, Gel	Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.	Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D005260	Female		Females