Unit

UNIT 5.4 Analysis of the Association of Proteins with Membranes

  1. T. Okamoto1,
  2. R.B. Schwab2,
  3. P.E. Scherer2,
  4. M.P. Lisanti2

Published Online: 1 MAY 2001

DOI: 10.1002/0471143030.cb0101s05

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Okamoto, T., Schwab, R., Scherer, P. and Lisanti, M. 2001. Analysis of the Association of Proteins with Membranes. Current Protocols in Cell Biology. 5:5.4:5.4.1–5.4.17.

Author Information

  1. 1

    Cleveland Clinic Foundation, Cleveland, Ohio

  2. 2

    Albert Einstein College of Medicine, Bronx, New York

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 2000

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.