caa a22 4500 397494483 CHVBK 20180308164517.0 cr unu---uuuuu 161202e199508 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a124915 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a124915 Comparison of Ca2+/Calmodulin-Dependent Protein Kinase IV from Rat Brain, Expressed in Insect Cells, and Expressed in Escherichia coli [Elektronische Daten] [Takako Kitani, Sachiko Okuno, Hitoshi Fujisawa] Calmodulin-dependent protein kinase IV (CaM-kinase IV) is thought to play crucial roles in the functioning of Ca2+ in the central nervous system and immune system, and the regulation of its activity is therefore very important. Recombinant CaM-kinase IV is invaluable for studies of its regulatory mechanism, because of its large-amount availability and ready site-specific mutagenesis. In the present study, rat CaM-kinase IV was expressed in Sf9 cells and Escherichia coli, and the kinetic properties were examined with syntide-2 and peptide-γ as substrates. The recombinant enzymes were produced highly efficiently, comprising as much as about 15% of the total protein in Sf9 cells and 9% in E.coli. The brain enzyme shows two Km values for syntide-2 in the presence of Ca2+/calmodulin, but the recombinant enzymes showed normal kinetic behavior. The brain enzyme and Sf9 enzyme showed Km values for peptide-γ of 53 and 82 μM, respectively, but the Km of the E. coli enzyme was as high as 1.7 mM, in the presence of Ca2+/calmodulin. Thus, the three enzymes differed in their kinetic properties, but all the three were markedly activated upon incubation with CaM-kinase IV kinase under the Ca2+/calmodulin-dependent protein phosphorylation conditions © by the Journal of Biochemistry Regular Papers nationallicence brain nationallicence Ca2+calmodulin nationallicence CaM-kinase IV nationallicence cDNA expression nationallicence protein kinase nationallicence Kitani Takako Affiliated with the Laboratory for Radioactive Isotope Research, Asahikawa Medical College aut Okuno Sachiko Department of Biochemistry, Asahikawa Medical College, Asahikawa, Hokkaido 078 aut Fujisawa Hitoshi Department of Biochemistry, Asahikawa Medical College, Asahikawa, Hokkaido 078 aut The Journal of Biochemistry Oxford University Press 118/2(1995-08), 364-370 0021-924X 118:2<364 1995 118 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a124915 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a124915 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kitani Takako Affiliated with the Laboratory for Radioactive Isotope Research, Asahikawa Medical College aut NATIONALLICENCE 700 1- Okuno Sachiko Department of Biochemistry, Asahikawa Medical College, Asahikawa, Hokkaido 078 aut NATIONALLICENCE 700 1- Fujisawa Hitoshi Department of Biochemistry, Asahikawa Medical College, Asahikawa, Hokkaido 078 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 118/2(1995-08), 364-370 0021-924X 118:2<364 1995 118 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