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Science 296 (5569): 913-916

Copyright © 2002 by the American Association for the Advancement of Science

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

David A. Zacharias,13*dagger Jonathan D. Violin,12* Alexandra C. Newton,1 Roger Y. Tsien13ddagger

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.

1 Department of Pharmacology,
2 Biomedical Sciences Graduate Program, and
3 Howard Hughes Medical Institute, University of California, San Diego, La Jolla, CA 92093-0647, USA.
*   These authors contributed equally to this work.

dagger    Present address: Merck Research Laboratories, 3535 General Atomics Court, MRLSDB1, San Diego, CA 92121, USA.

ddagger    To whom correspondence should be addressed. E-mail: rtsien{at}ucsd.edu

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K.-R. Lin, S.-F. Lee, C.-M. Hung, C.-L. Li, H.-F. Yang-Yen, and J. J. Y. Yen (2007)
J. Biol. Chem. 282, 21962-21972
   Abstract »    Full Text »    PDF »
FRET imaging in living maize cells reveals that plasma membrane aquaporins interact to regulate their subcellular localization.
E. Zelazny, J. W. Borst, M. Muylaert, H. Batoko, M. A. Hemminga, and F. Chaumont (2007)
PNAS 104, 12359-12364
   Abstract »    Full Text »    PDF »
Endothelial cytoskeletal reorganization in response to PAR1 stimulation is mediated by membrane rafts but not caveolae.
M. Carlile-Klusacek and V. Rizzo (2007)
Am J Physiol Heart Circ Physiol 293, H366-H375
   Abstract »    Full Text »    PDF »
Differential Association of Phosphatidylinositol 3-Kinase, SHIP-1, and PTEN with Forming Phagosomes.
L. A. Kamen, J. Levinsohn, and J. A. Swanson (2007)
Mol. Biol. Cell 18, 2463-2472
   Abstract »    Full Text »    PDF »
Signaling through a G Protein-coupled Receptor and Its Corresponding G Protein Follows a Stoichiometrically Limited Model.
F. Philip, P. Sengupta, and S. Scarlata (2007)
J. Biol. Chem. 282, 19203-19216
   Abstract »    Full Text »    PDF »
Receptor Complexes Cotransported via Polarized Endocytic Pathways Form Clusters with Distinct Organizations.
H. Wallrabe, G. Bonamy, A. Periasamy, and M. Barroso (2007)
Mol. Biol. Cell 18, 2226-2243
   Abstract »    Full Text »    PDF »
CD8 Raft Localization Is Induced by Its Assembly into CD8{alpha}beta Heterodimers, Not CD8{alpha}{alpha} Homodimers.
D. J. Pang, A. C. Hayday, and M.-J. Bijlmakers (2007)
J. Biol. Chem. 282, 13884-13894
   Abstract »    Full Text »    PDF »
Sterol-Rich Plasma Membrane Domains in Fungi.
F. J. Alvarez, L. M. Douglas, and J. B. Konopka (2007)
Eukaryot. Cell 6, 755-763
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Arabidopsis peroxin 16 trafficks through the ER and an intermediate compartment to pre-existing peroxisomes via overlapping molecular targeting signals.
S. K. Karnik and R. N. Trelease (2007)
J. Exp. Bot. 58, 1677-1693
   Abstract »    Full Text »    PDF »
R-Ras Regulates Exocytosis by Rgl2/Rlf-mediated Activation of RalA on Endosomes.
A. Takaya, T. Kamio, M. Masuda, N. Mochizuki, H. Sawa, M. Sato, K. Nagashima, A. Mizutani, A. Matsuno, E. Kiyokawa, et al. (2007)
Mol. Biol. Cell 18, 1850-1860
   Abstract »    Full Text »    PDF »
UvrD Limits the Number and Intensities of RecA-Green Fluorescent Protein Structures in Escherichia coli K-12.
R. C. Centore and S. J. Sandler (2007)
J. Bacteriol. 189, 2915-2920
   Abstract »    Full Text »    PDF »

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