AKAP350 at the Golgi Apparatus
AKAP350 can scaffold a number of protein kinases and phosphatases at the centrosome and the Golgi apparatus. We performed a yeast two-hybrid screen of a rabbit parietal cell library with a 3.2-kb segment of AKAP350 (nucleotides 3611â6813). This screen yielded a full-length clone of rabbit chloride...
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Published in | The Journal of biological chemistry Vol. 277; no. 43; pp. 40973 - 40980 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | English |
Published |
American Society for Biochemistry and Molecular Biology
25.10.2002
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Online Access | Get full text |
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Summary: | AKAP350 can scaffold a number of protein kinases and phosphatases at the centrosome and the Golgi apparatus. We performed
a yeast two-hybrid screen of a rabbit parietal cell library with a 3.2-kb segment of AKAP350 (nucleotides 3611â6813). This
screen yielded a full-length clone of rabbit chloride intracellular channel 1 (CLIC1). CLIC1 belongs to a family of proteins,
all of which contain a high degree of homology in their carboxyl termini. All CLIC family members were able to bind a 133-amino
acid domain within AKAP350 through the last 120 amino acids in the conserved CLIC carboxyl termini. Antibodies developed against
a bovine CLIC, p64, immunoprecipitated AKAP350 from HCA-7 colonic adenocarcinoma cell extracts. Antibodies against CLIC proteins
recognized at least five CLIC species including a novel 46-kDa CLIC protein. We isolated the human homologue of bovine p64,
CLIC5B, from HCA-7 cell cDNA. A splice variant of CLIC5, the predicted molecular mass of CLIC5B corresponds to the molecular
mass of the 46-kDa CLIC immunoreactive protein in HCA-7 cells. Antibodies against CLIC5B colocalized with AKAP350 at the Golgi
apparatus with minor staining of the centrosomes. AKAP350 and CLIC5B association with Golgi elements was lost following brefeldin
A treatment. Furthermore, GFP-CLIC5B-(178â410) targeted to the Golgi apparatus in HCA-7 cells. The results suggest that AKAP350
associates with CLIC proteins and specifically that CLIC5B interacts with AKAP350 at the Golgi apparatus in HCA-7 cells. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M112277200 |