A HIGH-AFFINITY BINDING-PROTEIN FOR THE REGULATORY SUBUNIT OF CAMP-DEPENDENT PROTEIN KINASE-II IN THE CENTROSOME OF HUMAN-CELLS

被引:64
作者
KERYER, G
RIOS, RM
LANDMARK, BF
SKALHEGG, B
LOHMANN, SM
BORNENS, M
机构
[1] UNIV OSLO, INST MED BIOCHEM, OSLO 3, NORWAY
[2] UNIV WURZBURG, DEPT MED, CLIN BIOCHEM LAB, W-8700 WURZBURG, GERMANY
关键词
D O I
10.1006/excr.1993.1029
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
In the human lymphoblastic cell line KE 37, Northern blot analysts with cDNA probes for human regulatory subunits RIIα and RIIβ of the cAMP-dependent protein kinaso (A-kinase) type II and immunoblotting or immunoprecipitation studies with several antibodies directed against RIIα and RIIβ show that these two isoforms are expressed. The major isoform α is mostly cytosolic, whereas the β isoform appears concentrated in the Golgi-centrosomal area, as judged by immunofluorescence and cell fractionation. Using a 32P-labeled 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. © 1993 Academic Press, Inc.
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页码:230 / 240
页数:11
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