Occurrence of Two Types of Ca²⁺‐Dependent Protein Kinases in the Cytosol Fraction of the Brain

Abstract

Two types of Ca2+-dependent protein kinases were demonstrated and partially purified from the cytosol fraction of rat brain by DEAE-cellulose, Sephadex G-200, and calmodulin-affinity column chromatography, using endogenous proteins and chicken gizzard myosin light chains as substrates. The molecular weights of the enzymes were 88,000 (peak I) and 120,000 (peak II) on gel filtration. Peak I had no affinity for calmodulin, whereas peak II had a high affinity for it, with a Ka value of 16.7 nM. The Ka values of peaks I and II for Ca2+ were 2.4 and 1.6 microM, respectively.

Autophosphorylation of Ca2+/calmodulin-dependent protein kinase. II. Effects on interaction between enzyme and substrate.

“…PSD themselves were used as the source of CaM kinase II. Using published methods, we prepared calmodulin-deficient MLC from chicken gizzard (20), MBP from bovine brain (21), synapsin I from rat brain (22), and tau factor and MAP 2 from porcine brain (23).…”

Ca2+, calmodulin‐dependent phosphorylation of glycogen synthase by a brain protein kinase

“…When indicated, the reaction was terminated by the addition of the SDS sample buffer [13]. Samples were subjected to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) in 9% acrylamide [13], followed by autoradiography [14]. The activity of glycogen synthase was determined in the presence of 10 mM glucose-6-phosphate as in [ 151.…”

Dissociation of alterations in levels of myosin light chain kinase and cyclic AMP-dependent protein kinase in rabbit myometrium during pregnancy.

“…Calmodulin was prepared from bovine brain according to the method of Yazawa et al (1980). Calmodulin-deficient myosin light chains from chicken gizzard myosin were obtained as described previously (Miyamoto et al, 1981a). All other chemicals were of reagent grade.…”

“…The assays for myosin light chain kinase and cyclic AMP-dependent protein kinase were performed by the methods of Miyamoto et al (1981a) Analysis of alteration in types I and II cyclic AMP-dependent protein kinase activities by DEAE-cellulose column chromatography during pregnancy:…”

“…The enzyme was also found in non-muscle tissues such as platelets (Daniel and Adelstein, 1976), brain (Dabrowska and Hartshorne, 1978;Yamauchi and Fujisawa, 1980;Hathaway et al, 1981;Miyamoto et al, 1981a;Fukunaga et al, 1982a;Fukunaga et al, 1982b) and other tissues (Miyamoto et al 1981b). The role of myosin light chain kinase in smooth muscle may relate to the contraction of the muscle (Gorecka et al, 1976;Adelstein and Eisenberg, 1980;Stull et al, 1980), since the phosphorylation of myosin dependent ATPase activity.…”