Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase.

Abstract

A calcium/calmodulin-dependent protein kinase type II (CaM-K) a-subunit cDNA has been cloned from rat brain. This enzyme is encoded by a 5.1-kilobase mRNA expressed exclusively in the brain. Hybridization histochemistry reveals that the CaM-K mRNA expression corresponds to the distribution of the immunoreactive a-subunit protein, suggesting that the high enzyme levels in specific brain areas reflect regional differences in gene expression. The sequence of CaM-K a-subunit cDNA indicates a 478-amino acid (54-kDa) protein with three functional domains. The domain organization suggests a structural model for calcium/calmodulindependent and independent states that might subserve shortand long-term responses to transient stimuli.

Calcium/Calmodulin-Dependent Protein Kinase II

“…Initially, this conclusion was based on the observations that, following separation of the subunits by SDS/polyacrylamide-gel electrophoresis and renaturation, both the a and fl subunits of the rat brain kinase bind 125-labelled calmodulin [7,11,12,14] and undergo autophosphorylation [43]. This conclusion has been confirmed by the recent publication of amino acid sequences for the a [31,32], , [44] and /1' [33] subunits of the rat brain kinase deduced from cDNA clones. All the rat brain kinase subunits contain the consensus sequences for ATP binding, protein kinase catalytic activity and calmodulinbinding domains, as depicted schematically in Fig.…”

“…Analysis of the amino acid sequence of CaM-kinase II for homologies with other calmodulin-binding proteins [31][32][33]44] indicates that the calmodulin-binding domain may be between residues 295 and 315 in the a-subunit. A synthetic peptide corresponding to this region interacts with calmodulin in a Ca2"-dependent manner [31].…”

Flattening the Glucocorticoid Rhythm causes Changes in Hippocampal Expression of Messenger RNAs Coding Structural and Functional Proteins: Implications for Aging and Depression

“…The MAP2b probe was a 39-mer (sequence 5 0 ctt cct cac ctc gaa cct ctg ctg gtt cgg aca caa ctg 3 0 ) targeted to base pairs 3781-3820 of the mRNA transcript (Marechal et al, 1988;Kindler et al, 1990). The probe for CAMKIIa was a 31-mer (5 0 cct ggt ccg gtc cca ggg acg cga gaa cga a 3 0 ) targeting base pairs 1480-1510 of the mRNA transcript (Lin et al, 1987). The probe for synaptophysin (5 0 tag cct tgc tgc cca tag tcg ccc tga ggc ccg tag cca 3 0 ) targeting base pairs 870-908 of the mRNA transcript (Bargou and Leube, 1991) was taken from the literature (Eastwood et al, 1994).…”

A non‐selective cation current activated via the multifunctional Ca(2+)‐calmodulin‐dependent protein kinase in human epithelial cells.

“…'Assoc' denotes the C-terminal association domain of the subunit. The numbers refer to the position of amino acids in the a-isozyme of CaM kinase (Lin et al 1987). Amino acid sequences of the peptides are abbreviated using their single letter code: A, alanine; C, cysteine; D, aspartic acid; E, glutamic acid; F, phenylalanine; G, glycine; H, histidine; I, isoleucine; K, lysine; L, leucine; M, methionine; N, asparagine; Q, glutamine; R, arginine; S, serine; T, threonine; V, valine.…”