Rapid cell surface appearance of endocytic membrane proteins in Chinese hamster ovary cells.

Abstract:

:Lactoperoxidase was used to selectively radiolabel endocytic membrane. CHO cells were incubated with enzyme at 37 degrees C for 10 min to permit lactoperoxidase internalization. Radioiodination was done at 4 degrees C. About 90% of the radioiodinated products pelleted at 100,000 X g. From 12 to 15 different electrophoretic species were detected by one-dimensional gel electrophoresis. When cells labeled by internalized lactoperoxidase were warmed to 37 degrees C, the incorporated radioactivity was lost in a biphasic manner with an overall t1/2 of approximately 20 h. Upon warming cells to 37 degrees C, the labeled species became sensitive to pronase or trypsin digestion. The increase in protease sensitivity was progressive over a 10- to 20-min period. Maximally 45% of the initially intracellular radiolabel could be released. A digest of exterior-radioiodinated cells released 50% of the incorporated radioiodine. These observations strongly suggest a rapid shuttling of approximately 90% of the radioiodinated membrane species initially present within the cell to the cell surface.

journal_name

Mol Cell Biol

authors

Storrie B,Dreesen TD,Maurey KM

doi

10.1128/mcb.1.3.261

subject

Has Abstract

pub_date

1981-03-01 00:00:00

pages

261-8

issue

3

eissn

0270-7306

issn

1098-5549

journal_volume

1

pub_type

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