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Degradation of proteins microinjected into cultured mammalian cells

✍ Scribed by Michael Zavortink; Thomas Thacher; Martin Rechsteiner

Publisher
John Wiley and Sons
Year
1979
Tongue
English
Weight
807 KB
Volume
100
Category
Article
ISSN
0021-9541

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✦ Synopsis

Abstract

Iodinated proteins were degraded after injection into HeLa cells at first‐order rates with half‐lives varying from three hours for the trout nonhistone chromosomal protein, HMG‐T, to 60 hours for whale myoglobin. Fluoresceinated‐bovine serum albumin (fl‐BSA) was degraded almost twice as fast as unmodified BSA. The rate of degradation of ^125^I‐BSA was very similar in eight cell lines of mouse, human, monkey and rat origin. Microinjected proteins were analyzed on SDS‐acrylamide gels after injection, and for BSA and immunoglobin G, all remaining intracellular ^125^I migrated at the molecular weight of the injected proteins. By contrast, more than 80% of the extracellular ^125^I chromatographed as iodotyrosine. With the exception of fl‐BSA, which exhibited perinuclear accumulation in approximately one‐half of the injected cells, autoradiography showed that throughout the period of study the injected proteins remained dispersed in the cytoplasm.

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