A conserved hydrophobic tetrad near the C terminus of the secretory Na+-K+-2Cl- cotransporter (NKCC1) is required for its correct intracellular processing.

The Journal of Biological Chemistry
Akihiro NezuR James Turner

Abstract

Little is known about the intracellular folding and trafficking of integral membrane proteins. Here we identify a hydrophobic amino acid tetrad (ILLV) close to the C terminus of the secretory Na+-K+-2Cl- cotransporter (NKCC1) that is important for the proper intracellular processing of this protein. This tetrad appears in a C-terminal sequence pattern that is conserved across species in a number of members of the NKCC1 gene family (slc12) of electroneutral salt transporters. We studied the effects of various mutations of these amino acids on NKCC1 transiently transfected into HEK-293 cells. Our results show that mutation of two of these residues to alanine leads to a >50% reduction in expression and complex glycosylation levels and that multiple mutations to alanine have cumulative effects. By contrast, scrambling of these amino acids, or mutation of other nearby conserved C-terminal residues, has little effect on these parameters. Mutation of ILLV to AAAA reduces complex glycosylation of NKCC1 by approximately 90% and results in a protein that does not form stable dimers and is retained in the endoplasmic reticulum in a highly aggregated state. Our results are consistent with the hypothesis that mutation of the hydrophobic tet...Continue Reading

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Citations

Oct 16, 2009·American Journal of Physiology. Cell Physiology·Qifei SunKelly G Ten Hagen
Sep 9, 2011·American Journal of Physiology. Renal Physiology·Gustavo R AresPablo A Ortiz
Nov 26, 2013·Molecular Biology and Evolution·Anna-Maria HartmannOlaf R P Bininda-Emonds
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Sep 10, 2015·International Journal of Cell Biology·Richa SinghMauricio Di Fulvio
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Oct 8, 2013·Pflügers Archiv : European journal of physiology·Nicolas Markadieu, Eric Delpire
Feb 11, 2020·Journal of Cellular Physiology·Salma OmerEric Delpire
Mar 22, 2013·American Journal of Physiology. Cell Physiology·Laura SmithCarole M Liedtke
Jan 31, 2021·International Journal of Molecular Sciences·Sunday Solomon JosiahJinwei Zhang
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