Helliwell, P.A., Rumsby, M.G. and Kellett, G.L. (2003) Intestinal Sugar Absorption is Regulated by Phosphorylation and Turnover of Protein Kinase C ߉I Mediated by Phosphatidylinositol 3-Kinase- and Mammalian Target of Rapamycin-dependent Pathways. Journal of Biological Chemistry, 278 (31). pp. 28644-28650. ISSN 0021-9258
Abstract
Stimulation of intestinal fructose absorption by phorbol 12-myristate 13-acetate (PMA) results from rapid insertion of GLUT2 into the brush-border membrane and correlates with protein kinase C (PKC) {beta}II activation. We have therefore investigated the role of phosphatidylinositol 3 (PI3)-kinase and mammalian target of rapamycin in the regulation of fructose absorption by PKC {beta}II phosphorylation. In isolated jejunal loops, stimulation of fructose absorption by PMA was inhibited by preperfusion with wortmannin or rapamycin, which blocked GLUT2 activation and insertion into the brush-border membrane. Antibodies to the last 18 and last 10 residues of the C-terminal region of PKC {beta}II recognized several species differentially in Western blots. Extensive cleavage of native enzyme (80/78 kDa) to a catalytic domain product of 49 kDa occurred. PMA and sugars provoked turnover and degradation of PKC {beta}II by dephosphorylation to a 42-kDa species, which was converted to polyubiquitylated species detected at 180 and 250+ kDa. PMA increased the level of the PKC {beta}II 49-kDa species, which correlates with the GLUT2 level; wortmannin and rapamycin blocked these effects of PMA. Rapamycin and wortmannin inhibited PKC {beta}II turnover. PI3-kinase, PDK-1, and protein kinase B were present in the brush-border membrane, where their levels were increased by PMA and blocked by the inhibitors. We conclude that GLUT2-mediated fructose absorption is regulated through PI3-kinase and mammalian target of rapamycin-dependent pathways, which control phosphorylation of PKC {beta}II and its substrate-induced turnover and ubiquitin-dependent degradation. These findings suggest possible mechanisms for short term control of intestinal sugar absorption by insulin and amino acids.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) |
Depositing User: | York RAE Import |
Date Deposited: | 23 Apr 2009 11:36 |
Last Modified: | 23 Apr 2009 11:36 |
Published Version: | http://dx.doi.org/10.1074/jbc.M301479200 |
Status: | Published |
Publisher: | The American Society for Biochemistry and Molecular Biology |
Identification Number: | 10.1074/jbc.M301479200 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:6600 |