RECONSTITUTION OF HETEROLOGOUS AND CHIMERIC CASEIN KINASE-II WITH RECOMBINANT SUBUNITS FROM HUMAN AND DROSOPHILA - IDENTIFICATION OF SPECIES-SPECIFIC DIFFERENCES IN THE BETA-SUBUNIT
Wj. Lin et al., RECONSTITUTION OF HETEROLOGOUS AND CHIMERIC CASEIN KINASE-II WITH RECOMBINANT SUBUNITS FROM HUMAN AND DROSOPHILA - IDENTIFICATION OF SPECIES-SPECIFIC DIFFERENCES IN THE BETA-SUBUNIT, Journal of protein chemistry, 13(2), 1994, pp. 217-225
Casein kinase IT is composed of two catalytic (alpha) and two regulato
ry (beta) subunits, the amino acid sequences of the alpha and beta sub
units are highly conserved between species. To examine whether heterol
ogous casein kinase II could be formed, recombinant alpha and beta sub
units from human and Drosophila were reconstituted from inclusion bodi
es. Casein kinase II containing either human alpha and Dposophila beta
or Drosophila alpha and human beta subunits exhibited enzymatic prope
rties similar to those of the homologous holoenzymes with regard to sp
ecific activity, salt optima, and autophosphorylation. However, renatu
ration and reconstitution of casein kinase II was dependent on the typ
e of beta subunits and the redox conditions, with the Drosophila beta
subunits requiring more reduced conditions. Chimeric beta subunits pre
pared from human and Drosophila cDNA revealed that the N-terminal regi
on was responsible for the requirement for the reduced redox state dur
ing renaturation. The N-terminal region also affected solubility and e
lectrophoretic mobility of the beta subunit.