PMID: 6411736Sep 1, 1983Paper

Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. II. Immunological quantitation

The Journal of Cell Biology
M Pesonen, K Simons

Abstract

The envelope of vesicular stomatitis virus was fused with the apical plasma membrane of Madin-Darby canine kidney cells by low pH treatment. The fate of the implanted G protein was then followed using a protein A-binding assay, which was designed to quantitate the amount of G protein in the apical and the basolateral membranes. The implanted G protein was rapidly internalized at 31 degrees C, whereas at 10 degrees C no uptake was observed. Already after 15 min at 31 degrees C, a fraction of the G protein could be detected at the basolateral membrane. After 60 min 25-48% of the G protein was basolateral as measured by the protein A-binding assay. At the same time, 25-33% of the implanted G protein was detected at the apical membrane. Internalization of G protein was not affected by 20 mM ammonium chloride or by 10 microM monensin. However, the endocytosed G protein accumulated in intracellular vacuoles and redistribution back to the plasma membrane was inhibited. We conclude that the implanted G protein was rapidly internalized from the apical surface of Madin-Darby canine kidney cells and a major fraction was routed to the basolateral domain.

References

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Citations

Nov 1, 1991·Journal of Cellular Biochemistry·E L MelbyK Sandvig
Jan 1, 1985·Pflügers Archiv : European journal of physiology·P J SalasE Rodriguez-Boulan
Jan 1, 1988·The Journal of Membrane Biology·G Lauer, W W Minuth
Apr 1, 1986·The Journal of Cell Biology·T A GottliebD D Sabatini
Dec 1, 1986·The Journal of Cell Biology·K S Matlin
Dec 5, 1985·Biochimica Et Biophysica Acta·S SpiegelJ S Handler
Feb 1, 1993·European Journal of Immunology·J MüllbergS Rose-John

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