title: Phospho-regulated ACAP4-Ezrin interaction is essential for histamine-stimulated parietal cell secretion.
abstract: The ezrin-radixin-moesin proteins provide a regulated linkage between membrane proteins and the cortical cytoskeleton and also participate in signal transduction pathways. Ezrin is localized to the apical membrane of parietal cells and couples the protein kinase A activation cascade to the regulated HCl secretion. Our recent proteomic study revealed a protein complex of ezrin-ACAP4-ARF6 essential for volatile membrane remodeling (Fang, Z., Miao, Y., Ding, X., Deng, H., Liu, S., Wang, F., Zhou, R., Watson, C., Fu, C., Hu, Q., Lillard, J. W., Jr., Powell, M., Chen, Y., Forte, J. G., and Yao, X. (2006) Mol. Cell Proteomics 5, 1437-1449). However, knowledge of whether ACAP4 physically interacts with ezrin and how their interaction is integrated into membrane-cytoskeletal remodeling has remained elusive. Here we provide the first evidence that ezrin interacts with ACAP4 in a protein kinase A-mediated phosphorylation-dependent manner through the N-terminal 400 amino acids of ACAP4. ACAP4 locates in the cytoplasmic membrane in resting parietal cells but translocates to the apical plasma membrane upon histamine stimulation. ACAP4 was precipitated with ezrin from secreting but not resting parietal cell lysates, suggesting a phospho-regulated interaction. Indeed, this interaction is abolished by phosphatase treatment and validated by an in vitro reconstitution assay using phospho-mimicking ezrin(S66D). Importantly, ezrin specifies the apical distribution of ACAP4 in secreting parietal cells because either suppression of ezrin or overexpression of non-phosphorylatable ezrin prevents the apical localization of ACAP4. In addition, overexpressing GTPase-activating protein-deficient ACAP4 results in an inhibition of apical membrane-cytoskeletal remodeling and gastric acid secretion. Taken together, these results define a novel molecular mechanism linking ACAP4-ezrin interaction to polarized epithelial secretion.
authors.affiliation: Anhui Key Laboratory of Cellular Dynamics and Chemical Biology, University of Science and Technology of China, Hefei 230027, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Ding
authors.forename: Hui
authors.initials: H
authors.lastname: Deng
authors.forename: Dongmei
authors.initials: D
authors.lastname: Wang
authors.forename: Jiajia
authors.initials: J
authors.lastname: Zhou
authors.forename: Yuejia
authors.initials: Y
authors.lastname: Huang
authors.forename: Xuannv
authors.initials: X
authors.lastname: Zhao
authors.forename: Xue
authors.initials: X
authors.lastname: Yu
authors.forename: Ming
authors.initials: M
authors.lastname: Wang
authors.forename: Fengsong
authors.initials: F
authors.lastname: Wang
authors.forename: Tarsha
authors.initials: T
authors.lastname: Ward
authors.forename: Felix
authors.initials: F
authors.lastname: Aikhionbare
authors.forename: Xuebiao
authors.initials: X
authors.lastname: Yao
mesh_terms: D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000378:metabolism; D000632:Aminopyrine / Q000737:chemistry; D000818:Animals; D001427:Bacterial Toxins / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000737:chemistry*; D004847:Epithelial Cells / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D006632:Histamine / Q000737:chemistry*; D010295:Parietal Cells, Gastric / Q000378:metabolism*; D010766:Phosphorylation; D006180:Proton-Translocating ATPases / Q000737:chemistry; D011817:Rabbits; D000099149:Ezrin
chemical_list: D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors; D000632:Aminopyrine; D001427:Bacterial Toxins; D017868:Cyclic AMP-Dependent Protein Kinases; D003598:Cytoskeletal Proteins; D020558:GTP Phosphohydrolases; D020690:GTPase-Activating Proteins; D006632:Histamine; D006180:Proton-Translocating ATPases; D000099149:Ezrin