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Atypon Free PMC Article

Mol Cell Biol. 1981 Mar;1(3):261-8. doi: 10.1128/mcb.1.3.261-268.1981.

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

Molecular and cellular biology

B Storrie, T D Dreesen, K M Maurey

Affiliations

  1. Department of Biochemistry and Nutrition, Virginia Polytechnic Institute and State University, Blacksburg 24061.

PMID: 6965100 PMCID: PMC369670 DOI: 10.1128/mcb.1.3.261-268.1981
Free PMC Article

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.

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