Journal
JOURNAL OF CELL BIOLOGY
Volume 165, Issue 5, Pages 735-746Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.200312170
Keywords
lipid rafts; membrane microdomains; lateral diffusion; fluorescence recovery after photobleaching; cholesterol
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Funding
- NCI NIH HHS [CA68485, P30 CA068485] Funding Source: Medline
- NIDDK NIH HHS [DK20593, P60 DK020593, P30 DK020593, P30 DK058404, DK58404] Funding Source: Medline
- Wellcome Trust Funding Source: Medline
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Lipid rafts are conceptualized as membrane microdomains enriched in cholesterol and glycosphingolipid that serve as platforms for protein segregation and signaling. The properties of these domains in vivo are unclear. Here, we use fluorescence recovery after photobleaching to test if raft association affects a protein's ability to laterally diffuse large distances across the cell surface. The diffusion coefficients (D) of several types of putative raft and nonraft proteins were systematically measured under steady-state conditions and in response to raft perturbations. Raft proteins diffused freely over large distances (>4 mum), exhibiting Ds that varied 10-fold. This finding indicates that raft proteins do not undergo long-range diffusion as part of discrete, stable raft domains. Perturbations reported to affect lipid rafts in model membrane systems or by biochemical fractionation (cholesterol depletion, decreased temperature, and cholesterol loading) had similar effects on the diffusional mobility of raft and nonraft proteins. Thus, raft association is not the dominant factor in determining long-range protein mobility at the cell surface.
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