En bloc incorporation of coatomer subunits during the assembly of COP- coated vesicles

S. Hara-Kuge, O. Kuge, L. Orci, M. Amherdt, M. Ravazzola, F. T. Wieland, J. E. Rothman

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Abstract

The cDNA encoding ε-COP, the 36-kD subunit of coatomer, was cloned from a bovine liver cDNA library and sequenced. Immunoblotting with an anti-ε-COP antibody showed that ε-COP exists in COP-coated vesicles as well as in the cytosolic coatomer. Using the cloned cDNA, recombinant His6-tagged ε-COP was overexpressed in cultured Chinese hamster ovary (CHO) cells, from which metabolically radiolabeled coatomer was purified by taking advantage of the His6 tag. Radiolabeled coatomer was employed to establish that all the subunits of the coatomer enter coated vesicles as an intact unit.

Original languageEnglish
Pages (from-to)883-892
Number of pages10
JournalJournal of Cell Biology
Volume124
Issue number6
DOIs
Publication statusPublished - 1994

All Science Journal Classification (ASJC) codes

  • Cell Biology

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    Hara-Kuge, S., Kuge, O., Orci, L., Amherdt, M., Ravazzola, M., Wieland, F. T., & Rothman, J. E. (1994). En bloc incorporation of coatomer subunits during the assembly of COP- coated vesicles. Journal of Cell Biology, 124(6), 883-892. https://doi.org/10.1083/jcb.124.6.883