Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells.
Many proteins associated with the plasma membrane are known to partition into submicroscopic sphingolipid- and cholesterol-rich domains called lipid rafts, but the determinants dictating this segregation of proteins in the membrane are poorly understood. We suppressed the tendency of Aequorea fluorescent proteins to dimerize and targeted these variants to the plasma membrane using several different types of lipid anchors. Fluorescence resonance energy transfer measurements in living cells revealed that acyl but not prenyl modifications promote clustering in lipid rafts. Thus the nature of the lipid anchor on a protein is sufficient to determine submicroscopic localization within the plasma membrane.
Zacharias
David A
DA
Department of Pharmacology, Biomedical Sciences Graduate Program, and, Howard Hughes Medical Institute, University of California, San Diego, La Jolla, CA 92093-0647, USA.
Violin
Jonathan D
JD
Newton
Alexandra C
AC
Tsien
Roger Y
RY
D000215:Acylation; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D051242:Caveolin 1; D022461:Caveolins / Q000378:metabolism; D002460:Cell Line; D003902:Detergents; D019281:Dimerization; D004285:Dogs; D004735:Energy Transfer; D005453:Fluorescence; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism*; D019814:Myristic Acid / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D019308:Palmitic Acid / Q000378:metabolism; D017368:Protein Prenylation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012995:Solubility; D013050:Spectrometry, Fluorescence; D014162:Transfection
D001426:Bacterial Proteins; D051242:Caveolin 1; D022461:Caveolins; D003902:Detergents; D008164:Luminescent Proteins; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D019814:Myristic Acid; D049452:Green Fluorescent Proteins; D019308:Palmitic Acid