Chloride transport in apical membrane vesicles from bovine tracheal epithelium: characterization using a fluorescent indicator.
C1 transport in apical membrane vesicles derived from bovine tracheal epithelial cells was studied using the C1-sensitive fluorescent indicator 6-methoxy-N-(3-sulfopropyl) quinolinium. With an inwardly directed 50 mM C1 gradient at 23 degrees C, the initial rate of C1 entry (JC1) was increased significantly from 0.32 +/- 0.12 nmol.sec-1.mg protein-1 (mean +/- SEM) to 0.50 +/- 0.07 nmol.sec-1.mg protein-1 when membrane potential was changed from 0 to +60 mV with K/valinomycin. At 37 degrees C, with membrane potential clamped at 0 mV, there was a 34 +/- 7% (n = 5) decrease in JC1 from a control value of 0.37 +/- 0.03 nmol.sec-1.mg protein-1 upon addition of 0.2 mM diphenylamine-2-carboxylate. The following did not alter JC1 significantly (JC1 values given as percent change from control): 50 mM cis Na (-1 +/- 5%), 0.1 mM furosemide (-3 +/- 4%), 0.1 mM furosemide in the presence of 50 mM cis Na (-5 +/- 2%), 0.1 mM H2DIDS (-18 +/- 9%), a 1.5 pH unit inwardly directed H gradient (-7 +/- 7%), and 0.1 mM H2DIDS in the presence of a 1.5 unit pH gradient (4 +/- 18%). With inward 50 mM anion gradients, the initial rates of Br and I entry (JBr and JI, respectively) were not significantly different from JC1.JC1 was a saturable function of C1 concentration with apparent Kd of 24 mM and apparent Vmax of 0.54 nmol.sec-1.mg protein-1.(ABSTRACT TRUNCATED AT 250 WORDS)
Fong
P
P
Department of Physiology, University of California, San Francisco.
Illsley
N P
NP
Widdicombe
J H
JH
Verkman
A S
AS
D000818:Animals; D001692:Biological Transport; D002417:Cattle; D002712:Chlorides / Q000378:metabolism / Q000493:pharmacokinetics*; D004848:Epithelium / Q000648:ultrastructure; D005456:Fluorescent Dyes; D007425:Intracellular Membranes / Q000502:physiology*; D015388:Organelles / Q000502:physiology*; D014132:Trachea / Q000166:cytology* / Q000648:ultrastructure
D002712:Chlorides; D005456:Fluorescent Dyes