DOI: 10.1126/science.1068539. Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live Cells

Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live Cells

10.1126/science.1068539

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

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.

Bibliography

Zacharias, D. A., Violin, J. D., Newton, A. C., & Tsien, R. Y. (2002). Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live Cells. Science, 296(5569), 913â916.

Authors 4

David A. Zacharias (first)
Jonathan D. Violin (additional)
Alexandra C. Newton (additional)
Roger Y. Tsien (additional)

References 25 Referenced 1,998

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CFP and YFP were enhanced versions ECFP and YFP 10C Q69K (9). The signal for geranylgeranylation (GerGer) was a COOH-terminal CLLL from Rho; PalmPalm an NH 2 -terminal MLCCMRRTKQ from Gap43; MyrPalm an NH 2 -terminal MGCIKSKRKDNLNDDE from Lyn kinase. Full-length cDNA of bovine caveolin-1 was fused to the NH 2 -terminus of CFP and YFP. Single-letter abbreviations for the amino acid residues are as follows: A Ala; C Cys; D Asp; E Glu; F Phe; G Gly; H His; I Ile; K Lys; L Leu; M Met; N Asn; P Pro; Q Gln; R Arg; S Ser; T Thr; V Val; W Trp; and Y Tyr.
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We thank L. Burns H. Purkey and M. Petrassi for help with analytical ultracentrifugation and S. Adams G. Walkup R. Wachter and J. Henley for discussion and suggestions. Supported by NIH grants NS27177 (R.T.) DK54441 (A.N.) 2T32 GM07752 (J.V.) and HHMI.

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:54 a.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 9:22 p.m.)
Indexed	1 week, 1 day ago (Aug. 19, 2025, 6:52 a.m.)
Issued	23 years, 3 months ago (May 3, 2002)
Published	23 years, 3 months ago (May 3, 2002)
Published Print	23 years, 3 months ago (May 3, 2002)

Funders 0

None

BibTeX

@article{Zacharias_2002, title={Partitioning of Lipid-Modified Monomeric GFPs into Membrane Microdomains of Live Cells}, volume={296}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1068539}, DOI={10.1126/science.1068539}, number={5569}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Zacharias, David A. and Violin, Jonathan D. and Newton, Alexandra C. and Tsien, Roger Y.}, year={2002}, month=may, pages={913–916} }

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