title: An inactive mutant of the alpha subunit of protein kinase CK2 that traps the regulatory CK2beta subunit.
abstract: Protein kinase CK2 (casein kinase 2) is a ubiquitous Ser/Thr protein kinase involved in cell proliferation. Mutation of the alpha subunit of the Xenopus laevis CK2 to change aspartic acid 156 to alanine (CK2alphaA156) resulted in an inactive enzyme. The CK2alphaA156 mutant, however, binds the regulatory subunit as measured by retention of beta on a nickel chelating column mediated by (His)6-tagged CK2alphaA156. Addition of CK2alphaA156 also caused beta to shift sedimentation in a sucrose gradient from a beta2 dimer (52 kDa) to an alpha2beta2 tetramer (130,000 kDa). CK2alphaA156 can trap the beta subunit in an inactive complex reducing the stimulation of casein phosphorylation caused by addition of beta to wild-type alpha. This competitive effect depends on the ratio of alpha/alphaA156 and on the amount of beta available. Since beta inhibits the phosphorylation of calmodulin by CK2alpha, the addition of CK2alphaA156, in this case, increases calmodulin phosphorylation by the alpha and beta combination. These results suggest that CK2alphaA156 may be a useful dominant-negative mutant that can serve to explore the multiple functions of CK2beta.
authors.affiliation: Programa de Biologia Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago.
authors.forename: D
authors.initials: D
authors.lastname: Cosmelli
authors.forename: M
authors.initials: M
authors.lastname: Antonelli
authors.forename: C C
authors.initials: CC
authors.lastname: Allende
authors.forename: J E
authors.initials: JE
authors.lastname: Allende
mesh_terms: D000409:Alanine / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001224:Aspartic Acid / Q000737:chemistry / Q000378:metabolism; D001667:Binding, Competitive; D047390:Casein Kinase II; D016297:Mutagenesis, Site-Directed; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D014982:Xenopus laevis
chemical_list: D011993:Recombinant Fusion Proteins; D001224:Aspartic Acid; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D000409:Alanine