Cyclic AMP-mediated endocytosis of intestinal epithelial NHE3 requires binding to synaptotagmin 1.

American Journal of Physiology. Gastrointestinal and Liver Physiology
Mark W MuschEugene B Chang

Abstract

The apical membrane Na(+)-H(+) exchanger (NHE)3 is regulated by cAMP-dependent phosphorylation, which inhibits its activity through membrane endocytosis. The clathrin complex adaptor protein synaptotagmin 1 (Syt 1) appears to be essential to this process, but little is known about its expression in intestinal epithelial cells or interaction with NHE3. The intestinal epithelial expression and apical location of Syt 1 were determined by Syt 1 mRNA profiling and immunolocalization. Tandem mass spectrometry was used for protein identification. Bis(sulfosuccinimidyl) suberate (BS(3)) cross linking suggested that NHE3 and Syt 1 were in a membrane complex following cAMP stimulation of Caco2BBE (Brush Border Expressions) cells. To investigate the regulation of NHE3 appearance in a Syt 1-containing membrane compartment, doxycycline-inducible hemaglutinin (HA)-tagged NHE3 was expressed in Caco2BBE cells. HA-NHE3 correctly targeted to the apical membrane, where, upon cAMP stimulation, it was internalized with a Syt 1-containing compartment. Site-directed mutagenesis of NHE3 showed that serine 605 (S605) was pivotal to NHE3 and Syt 1 association and internalization. Direct Syt 1 interaction with NHE3 was suggested by fluorescence resonance...Continue Reading

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Citations

Jun 4, 2011·American Journal of Physiology. Gastrointestinal and Liver Physiology·Michelle A FalkowskiGuy E Groblewski
Sep 1, 2012·American Journal of Physiology. Gastrointestinal and Liver Physiology·Robert L JakabNadia A Ameen
Jul 19, 2012·World Journal of Gastroenterology : WJG·Alan B R ThomsonHugh Freeman
Nov 9, 2010·Annual Review of Physiology·Akira Kato, Michael F Romero
Nov 2, 2016·Journal of the American Society of Nephrology : JASN·Wilna OosthuyzenJames W Dear
Mar 5, 2015·Gene·Sara Sancho-KnapikJesús Osada
Aug 30, 2013·American Journal of Physiology. Cell Physiology·Anne M CollacoNadia A Ameen
Oct 9, 2015·American Journal of Physiology. Cell Physiology·Tarunmeet GujralPradeep K Dudeja
Dec 31, 2020·Journal of Clinical Medicine·Changsen LengSven C D van IJzendoorn

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