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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}

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J. Neurosci. 31, 11231-11243
   Abstract »    Full Text »    PDF »
Identification of an Anchor Residue for CheA-CheY Interactions in the Chemotaxis System of Escherichia coli.
H. Thakor, S. Nicholas, I. M. Porter, N. Hand, and R. C. Stewart (2011)
J. Bacteriol. 193, 3894-3903
   Abstract »    Full Text »    PDF »
Prenylated C17orf37 Induces Filopodia Formation to Promote Cell Migration and Metastasis.
S. Dasgupta, I. Cushman, M. Kpetemey, P. J. Casey, and J. K. Vishwanatha (2011)
J. Biol. Chem. 286, 25935-25946
   Abstract »    Full Text »    PDF »
Functional Cooperation between the Proteins Nck and ADAP Is Fundamental for Actin Reorganization.
M. H. Pauker, B. Reicher, S. Fried, O. Perl, and M. Barda-Saad (2011)
Mol. Cell. Biol. 31, 2653-2666
   Abstract »    Full Text »    PDF »
An Ion-insensitive cAMP Biosensor for Long Term Quantitative Ratiometric Fluorescence Resonance Energy Transfer (FRET) Measurements under Variable Physiological Conditions.
P. S. Salonikidis, M. Niebert, T. Ullrich, G. Bao, A. Zeug, and D. W. Richter (2011)
J. Biol. Chem. 286, 23419-23431
   Abstract »    Full Text »    PDF »
Symbiotic Rhizobia Bacteria Trigger a Change in Localization and Dynamics of the Medicago truncatula Receptor Kinase LYK3.
C. H. Haney, B. K. Riely, D. M. Tricoli, D. R. Cook, D. W. Ehrhardt, and S. R. Long (2011)
PLANT CELL 23, 2774-2787
   Abstract »    Full Text »    PDF »
AP-1 and clathrin are essential for secretory granule biogenesis in Drosophila.
J. Burgess, M. Jauregui, J. Tan, J. Rollins, S. Lallet, P. A. Leventis, G. L. Boulianne, H. C. Chang, R. Le Borgne, H. Kramer, et al. (2011)
Mol. Biol. Cell 22, 2094-2105
   Abstract »    Full Text »    PDF »
Direct Membrane Association Drives Mitochondrial Fission by the Parkinson Disease-associated Protein {alpha}-Synuclein.
K. Nakamura, V. M. Nemani, F. Azarbal, G. Skibinski, J. M. Levy, K. Egami, L. Munishkina, J. Zhang, B. Gardner, J. Wakabayashi, et al. (2011)
J. Biol. Chem. 286, 20710-20726
   Abstract »    Full Text »    PDF »
Development of a high-dynamic range, GFP-based FRET probe sensitive to oxidative microenvironments.
V. L. Kolossov, B. Q. Spring, R. M. Clegg, J. J. Henry, A. Sokolowski, P. J. A. Kenis, and H. R. Gaskins (2011)
Experimental Biology and Medicine 236, 681-691
   Abstract »    Full Text »    PDF »
FERM Domain Phosphoinositide Binding Targets Merlin to the Membrane and Is Essential for Its Growth-Suppressive Function.
T. Mani, R. F. Hennigan, L. A. Foster, D. G. Conrady, A. B. Herr, and W. Ip (2011)
Mol. Cell. Biol. 31, 1983-1996
   Abstract »    Full Text »    PDF »
The Serotonin Transporter Is an Exclusive Client of the Coat Protein Complex II (COPII) Component SEC24C.
S. Sucic, A. El-Kasaby, O. Kudlacek, S. Sarker, H. H. Sitte, P. Marin, and M. Freissmuth (2011)
J. Biol. Chem. 286, 16482-16490
   Abstract »    Full Text »    PDF »
Relationship between Homo-oligomerization of a Mammalian Olfactory Receptor and Its Activation State Demonstrated by Bioluminescence Resonance Energy Transfer.
F. Wade, A. Espagne, M.-A. Persuy, J. Vidic, R. Monnerie, F. Merola, E. Pajot-Augy, and G. Sanz (2011)
J. Biol. Chem. 286, 15252-15259
   Abstract »    Full Text »    PDF »
Revealing protein oligomerization and densities in situ using spatial intensity distribution analysis.
A. G. Godin, S. Costantino, L.-E. Lorenzo, J. L. Swift, M. Sergeev, A. Ribeiro-da-Silva, Y. De Koninck, and P. W. Wiseman (2011)
PNAS 108, 7010-7015
   Abstract »    Full Text »    PDF »
Protein Targeting to Exosomes/Microvesicles by Plasma Membrane Anchors.
B. Shen, N. Wu, J.-M. Yang, and S. J. Gould (2011)
J. Biol. Chem. 286, 14383-14395
   Abstract »    Full Text »    PDF »

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