Analysis of the association of proteins with membranes.

T. Okamoto, R. B. Schwab, P. E. Scherer, M. P. Lisanti

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Abstract

Proteins associate with membranes in a number of ways. Is the membrane an integral membrane protein? A peripheral protein? Does it have a glycosylphosphoinositol (GPI) tail? Is it associated with caveolae-derived plasma membrane domains? This unit describes methods for determining how a protein of interest associates with the membrane including extraction with alkaline carbonate, urea, high salt, or Triton X-114. GPI linkage is probed using PI-PLC cleavage. There is a method for detergent solubilization of Triton-X 100 insoluble integral and GPI-linked proteins. Caveolae membranes can be isolated using Triton X-100 or a detergent-free method.

Original languageEnglish (US)
JournalCurrent protocols in cell biology / editorial board, Juan S. Bonifacino ... [et al.]
VolumeChapter 5
Publication statusPublished - May 2001

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ASJC Scopus subject areas

  • Medicine(all)

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