caa a22 4500 397555237 CHVBK 20180308164759.0 cr unu---uuuuu 161202e199602 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021234 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021234 Purification and Characterization of Active Fragment of Ca2+Calmodulin-Dependent Protein Kinase II from the Post-Synaptic Density in the Rat Forebrain [Elektronische Daten] [Yoshiyuki Yoshimura, Tomohiko Nomura, Takashi Yamauchi] Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) of the post-synaptic density (PSD) was solubilized and activated 4- to 5-fold by limited α-chymotrypsin digestion with prior autophosphorylation of the kinase. The enzyme was also activated by trypsin and μ-calpain, a Ca2+-dependent protease. The active catalytic fragment was purified to homogeneity using gel filtration and ion exchange chromatography. The purified active fragment was completely Ca2+/calmodulin-independent and exists as a monomer. Kinetic studies with the purified fragment revealed similar Km values for ATP and synthetic peptide substrate, and an about 8-fold increment in Vmax, compared with native PSD CaM kinase II. © by the Journal of Biochemistry Regular Papers nationallicence active catalytic fragment nationallicence Ca2+/calmodulin-dependent protein kinase II nationallicence post-synaptic density nationallicence protein phosphorylation nationallicence rat forebrain nationallicence Yoshimura Yoshiyuki Department of Biochemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770 aut Nomura Tomohiko Department of Biochemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770 aut Yamauchi Takashi Department of Biochemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770 aut The Journal of Biochemistry Oxford University Press 119/2(1996-02), 268-273 0021-924X 119:2<268 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021234 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021234 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yoshimura Yoshiyuki Department of Biochemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770 aut NATIONALLICENCE 700 1- Nomura Tomohiko Department of Biochemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770 aut NATIONALLICENCE 700 1- Yamauchi Takashi Department of Biochemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/2(1996-02), 268-273 0021-924X 119:2<268 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford