Regulation of the amiloride-sensitive epithelial sodium channel by syntaxin 1A.
The first step in transepithelial sodium absorption lies at the apical membrane where the amiloride-sensitive, epithelial sodium channel, ENaC, facilitates sodium entry into the cell. Here we report that the vesicle traffic regulatory (SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)) protein, syntaxin 1A (S1A), inhibits ENaC mediated sodium entry. This inhibitory effect is selective for S1A and is not reproduced by syntaxin 3. The inhibition does not require the membrane anchoring domain of syntaxin 1A. It was reversed by the S1A-binding protein, Munc-18, but not by a Munc-18 mutant, which lacks syntaxin affinity. Immunostaining of epitope-tagged ENaC subunits showed that syntaxin 1A decreases ENaC current by reducing the number of ENaC channels in the plasma membrane; S1A does not interfere with ENaC protein expression. Immunoprecipitation of syntaxin 1A from the sodium-transporting epithelial cell line, A6, co-precipitates ENaC. These findings indicate that syntaxin 1A and other members of the SNARE machinery are involved in the control of plasma membrane ENaC content, and they suggest that SNARE proteins participate in the regulation of sodium absorption in relation to agonist mediated vesicle insertion-retrieval processes.
Qi
J
J
Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261, USA.
Peters
K W
KW
Liu
C
C
Wang
J M
JM
Edinger
R S
RS
Johnson
J P
JP
Watkins
S C
SC
Frizzell
R A
RA
D000818:Animals; D000954:Antigens, Surface / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D053503:Epithelial Sodium Channels; D006801:Humans; D051938:Munc18 Proteins; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D009842:Oligopeptides / Q000096:biosynthesis; D009865:Oocytes / Q000502:physiology; D010455:Peptides / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels / Q000235:genetics / Q000502:physiology*; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins
D000954:Antigens, Surface; D053503:Epithelial Sodium Channels; D051938:Munc18 Proteins; D009419:Nerve Tissue Proteins; D009842:Oligopeptides; D010455:Peptides; D011994:Recombinant Proteins; C494212:STX1A protein, human; D015222:Sodium Channels; D050827:Syntaxin 1; D033921:Vesicular Transport Proteins; C085706:FLAG peptide; D012964:Sodium