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J Biol Chem, Vol. 275, Issue 20, 15279-15286, May 19, 2000
From the Lipids and other membrane constituents recycle
between the plasma membrane and intracellular endocytic compartments.
In CHO cells, approximately half of the internalized
C6-NBD-SM, a fluorescent lipid analogue widely used
as a membrane maker, recycles via the endocytic recycling compartment
with a t1/2 of ~12 min (Mayor, S., Presley,
J. F., and Maxfield, F. R. (1993) J. Cell
Biol. 121, 1257-1269). Surprisingly, the rest returns to the
plasma membrane very quickly. A detailed kinetic study presented in
this paper indicates that after a brief internalization pulse, 42-62%
of the internalized C6-NBD-SM returns to the plasma membrane with a t1/2 of 1-2 min. Similar results
are obtained using HEp2 and nonpolarized Madin-Darby canine kidney
cells. Using FM dyes of different hydrophobicity, we show that rapid
recycling involves passage through an endocytic organelle that was
subsequently identified as the sorting endosome by co-localization with
internalized transferrin and low density lipoprotein. These results
imply that the membrane internalization rate is much higher than
previously estimated, with a t1/2 as short as 5-10
min. Rapid internalization and recycling would facilitate processes
such as nutrient uptake and cholesterol efflux.
Characterization of Rapid Membrane Internalization and
Recycling*
§ and¶
Department of Biochemistry, Weill Medical
College of Cornell University, New York, New York 10021 and the
§ Department of Chemistry and Chemical Biology, Cornell
University, Ithaca, New York 14853
*
This work was supported by National Institutes of Health
Grant DK 27083.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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