Distribution of A-kinase anchoring proteins in parietal cells.
Recent investigations have suggested that subcellular compartmentalization of second messenger responsive enzyme systems may be responsible for specific patterns of cellular activation. The type II cAMP-dependent kinase (A-kinase) is localized to particular subcellular domains through the binding of the regulatory subunit (RII) dimer to A-kinase anchoring proteins (AKAPs). Using a [32P]RII overlay assay, we have investigated the presence of AKAPs throughout the gastrointestinal tract, with specific emphasis focused on the gastric parietal cell. All gastrointestinal tissues contained at least one detectable AKAP (60 kDa), with five AKAPs (50-140 kDa) in fundic and antral mucosa. Isolated gastric glands contained four AKAPs. Two AKAPs (50 and 78 kDa) were detected in purified parietal cells, with the 78 kDa AKAP (AKAP78) specific to parietal cell enriched populations. RII-binding to all of these AKAPs was abolished by preincubation of [32P]RII with a synthetic peptide representing the RII-binding region of the AKAP, HT-31. AKAP78 was distributed throughout all membrane fractions of subfractionated parietal cells, with the largest amount of RII-binding detected in the light membrane fraction. Identification of A-kinase regulatory subunits by photoaffinity labeling with 8-azido-[32P]cAMP demonstrated that RII segregated into the same parietal cell subfractions as AKAP78. A majority (approximately 60%) of AKAP78 was detected in the Triton X-100-insoluble fraction, suggesting that this protein resides in a cytoskeletal domain. AKAP78 may be involved in localizing the type II A-kinase to specific intracellular locations in the parietal cell.
D000818:Animals; D002451:Cell Compartmentation; D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D005753:Gastric Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D008970:Molecular Weight; D010750:Phosphoproteins / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011817:Rabbits; D015398:Signal Transduction; D000099149:Ezrin
D054746:Cyclic AMP-Dependent Protein Kinase Type II; D017868:Cyclic AMP-Dependent Protein Kinases; D003598:Cytoskeletal Proteins; D010750:Phosphoproteins; D011506:Proteins; D000099149:Ezrin