title: A high-affinity binding protein for the regulatory subunit of cAMP-dependent protein kinase II in the centrosome of human cells.
abstract: In the human lymphoblastic cell line KE 37, Northern blot analysis with cDNA probes for human regulatory subunits RII alpha RII beta of the cAMP-dependent protein kinase (A-kinase) type II and immunoblotting or immunoprecipitation studies with several antibodies directed against RII alpha and RII beta show that these two isoforms are expressed. The major isoform alpha is mostly cytosolic, whereas the beta isoform appears concentrated in the Golgi-centrosomal area, as judged by immunofluorescence and cell fractionation. Using a 32P-labelled RII overlay on Western blots, a 350-kDa RII-binding protein (AKAP 350) was specifically identified in centrosomes isolated from this cell line, whereas a Golgi fraction has previously been demonstrated to contain an 85-kDa RII-binding protein (AKAP 85). AKAP 350 is highly insoluble and can partially be extracted from centrosomes as a complex of AKAP 350 and RII subunit. AKAP 350 was identified as a specific centrosomal protein previously demonstrated in the pericentriolar material. The potential significance of a specific subcellular distribution for different RII-binding proteins in nonneuronal cells is discussed.
authors.affiliation: Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, Gif/Yvette, France.
authors.forename: G
authors.initials: G
authors.lastname: Keryer
authors.forename: R M
authors.initials: RM
authors.lastname: Rios
authors.forename: B F
authors.initials: BF
authors.lastname: Landmark
authors.forename: B
authors.initials: B
authors.lastname: Skalhegg
authors.forename: S M
authors.initials: SM
authors.lastname: Lohmann
authors.forename: M
authors.initials: M
authors.lastname: Bornens
mesh_terms: D002460:Cell Line; D002502:Centrioles / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D011506:Proteins / Q000032:analysis*
chemical_list: D011506:Proteins; D011494:Protein Kinases