title: Reconstitution of heterologous and chimeric casein kinase II with recombinant subunits from human and Drosophila: identification of species-specific differences in the beta subunit.
abstract: Casein kinase II is composed of two catalytic (alpha) and two regulatory (beta) subunits, the amino acid sequences of the alpha and beta subunits are highly conserved between species. To examine whether heterologous casein kinase II could be formed, recombinant alpha and beta subunits from human and Drosophila were reconstituted from inclusion bodies. Casein kinase II containing either human alpha and Drosophila beta or Drosophila alpha and human beta subunits exhibited enzymatic properties similar to those of the homologous holoenzymes with regard to specific activity, salt optima, and autophosphorylation. However, renaturation and reconstitution of casein kinase II was dependent on the type of beta subunits and the redox conditions, with the Drosophila beta subunits requiring more reduced conditions. Chimeric beta subunits prepared from human and Drosophila cDNA revealed that the N-terminal region was responsible for the requirement for the reduced redox state during renaturation. The N-terminal region also affected solubility and electrophoretic mobility of the beta subunit.
authors.affiliation: Department of Biochemistry, University of California, Riverside 92521.
authors.forename: W J
authors.initials: WJ
authors.lastname: Lin
authors.forename: R
authors.initials: R
authors.lastname: Jakobi
authors.forename: J A
authors.initials: JA
authors.lastname: Traugh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D047390:Casein Kinase II; D003001:Cloning, Molecular; D004330:Drosophila / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D013045:Species Specificity
chemical_list: D011993:Recombinant Fusion Proteins; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases