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Development and Use of Fluorescent Protein Markers in Living Cells
Jennifer Lippincott-Schwartz,*George H. Patterson
The ability to visualize, track, and quantify molecules and
events in living cells with high spatial and temporal resolutionis
essential for understanding biological systems. Only recentlyhas it
become feasible to carry out these tasks due to the adventof
fluorescent protein technology. Here, we trace the developmentof
highly visible and minimally perturbing fluorescent proteinsthat,
together with updated fluorescent imaging techniques, areproviding
unprecedented insights into the movement of proteinsand their
interactions with cellular components in living cells.
Cell Biology and Metabolism Branch, National Institute of Child
Health and Human Development, National Institutes of Health, Bethesda,
MD 20892, USA.
*
To whom correspondence should be addressed. E-mail:
jlippin{at}helix.nih.gov
Daniel Zicha, Ian M. Dobbie, Mark R. Holt, James Monypenny, Daniel Y. H. Soong, Colin Gray, and Graham A. Dunn (4 April 2003) Science300 (5616), 142.
[DOI: 10.1126/science.1082026] |Abstract »|Full Text »|PDF »|Supporting Online Material »
PERSPECTIVES
Paul Martin and Susan M. Parkhurst (4 April 2003) Science300 (5616), 63.
[DOI: 10.1126/science.1084148] |Summary »|Full Text »|PDF »
INTRODUCTION TO SPECIAL ISSUE
Stella M. Hurtley and Laura Helmuth (4 April 2003) Science300 (5616), 75.
[DOI: 10.1126/science.300.5616.75] |Summary »|PDF »
NEWS
Mary Beckman (4 April 2003) Science300 (5616), 76.
[DOI: 10.1126/science.300.5616.76] |Summary »|Full Text »|PDF »
NEWS
Greg Miller (4 April 2003) Science300 (5616), 78.
[DOI: 10.1126/science.300.5616.78] |Summary »|Full Text »|PDF »
NEWS
Caroline Seydel (4 April 2003) Science300 (5616), 80.
[DOI: 10.1126/science.300.5616.80] |Summary »|Full Text »|PDF »
Novel Internal Regions of Fluorescent Proteins Undergo Divergent Evolutionary Patterns.
D. F. Gruber, R. DeSalle, E. K. Lienau, D. Tchernov, V. A. Pieribone, and H.-T. Kao (2009)
Mol. Biol. Evol.
26, 2841-2848
|Abstract »|Full Text »|PDF »
Unifying Fluorescence Microscopy and Mass Spectrometry for Studying Protein Complexes in Cells.
C. Deng, X. Xiong, and A. N. Krutchinsky (2009)
Mol. Cell. Proteomics
8, 1413-1423
|Abstract »|Full Text »|PDF »
Engineering of a monomeric fluorescent protein AsGFP499 and its applications in a dual translocation and transcription assay.
A. Tasdemir, F. Khan, T. A. Jowitt, L. Iuzzolino, S. Lohmer, S. Corazza, and T. J. Schmidt (2008)
Protein Eng. Des. Sel.
21, 613-622
|Abstract »|Full Text »|PDF »
Surface Trafficking of Neurotransmitter Receptor: Comparison between Single-Molecule/Quantum Dot Strategies.
L. Groc, M. Lafourcade, M. Heine, M. Renner, V. Racine, J.-B. Sibarita, B. Lounis, D. Choquet, and L. Cognet (2007)
J. Neurosci.
27, 12433-12437
|Full Text »|PDF »
Ligand-Dependent Reduction in the Membrane Mobility of FLAGELLIN SENSITIVE2, an Arabidopsis Receptor-Like Kinase.
G. S. Ali, K. V. S. K. Prasad, I. Day, and A. S. N. Reddy (2007)
Plant Cell Physiol.
48, 1601-1611
|Abstract »|Full Text »|PDF »
Nuclear Actin Regulates Dynamic Subcellular Localization and Activity of the SRF Cofactor MAL.
M. K. Vartiainen, S. Guettler, B. Larijani, and R. Treisman (2007)
Science
316, 1749-1752
|Abstract »|Full Text »|PDF »
Subcellular location of serum- and glucocorticoid-induced kinase-1 in renal and mammary epithelial cells.
E. Cordas, A. Naray-Fejes-Toth, and G. Fejes-Toth (2007)
Am J Physiol Cell Physiol
292, C1971-C1981
|Abstract »|Full Text »|PDF »
Differential dynamics of Rab3A and Rab27A on secretory granules.
M. T. W. Handley, L. P. Haynes, and R. D. Burgoyne (2007)
J. Cell Sci.
120, 973-984
|Abstract »|Full Text »|PDF »
Characterization and application of single fluorescent nanodiamonds as cellular biomarkers.
C.-C. Fu, H.-Y. Lee, K. Chen, T.-S. Lim, H.-Y. Wu, P.-K. Lin, P.-K. Wei, P.-H. Tsao, H.-C. Chang, and W. Fann (2007)
PNAS
104, 727-732
|Abstract »|Full Text »|PDF »
Plant G protein heterotrimers require dual lipidation motifs of G{alpha} and G{gamma} and do not dissociate upon activation.
M. J. W. Adjobo-Hermans, J. Goedhart, and T. W. J. Gadella Jr (2006)
J. Cell Sci.
119, 5087-5097
|Abstract »|Full Text »|PDF »
Breaking the resolution limit in light microscopy.
Depletion of type IA regulatory subunit (RI{alpha}) of protein kinase A (PKA) in mammalian cells and tissues activates mTOR and causes autophagic deficiency.
M. Mavrakis, J. Lippincott-Schwartz, C. A. Stratakis, and I. Bossis (2006)
Hum. Mol. Genet.
15, 2962-2971
|Abstract »|Full Text »|PDF »
Imaging Intracellular Fluorescent Proteins at Nanometer Resolution.
E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess (2006)
Science
313, 1642-1645
|Abstract »|Full Text »|PDF »
Polyglutamine proteins at the pathogenic threshold display neuron-specific aggregation in a pan-neuronal Caenorhabditis elegans model..
H. R. Brignull, F. E. Moore, S. J. Tang, and R. I. Morimoto (2006)
J. Neurosci.
26, 7597-7606
|Abstract »|Full Text »|PDF »
Visualization of intracellular transport of vesicular stomatitis virus nucleocapsids in living cells..
S. C. Das, D. Nayak, Y. Zhou, and A. K. Pattnaik (2006)
J. Virol.
80, 6368-6377
|Abstract »|Full Text »|PDF »
The vacuole system is a significant intracellular pathway for longitudinal solute transport in basidiomycete fungi..
P. R. Darrah, M. Tlalka, A. Ashford, S. C. Watkinson, and M. D. Fricker (2006)
Eukaryot. Cell
5, 1111-1125
|Abstract »|Full Text »|PDF »
The secretory membrane system in the Drosophila syncytial blastoderm embryo exists as functionally compartmentalized units around individual nuclei.
D. Frescas, M. Mavrakis, H. Lorenz, R. DeLotto, and J. Lippincott-Schwartz (2006)
J. Cell Biol.
173, 219-230
|Abstract »|Full Text »|PDF »
Messages diffuse faster than messengers.
B. Pando, S. P. Dawson, D.-O. D. Mak, and J. E. Pearson (2006)
PNAS
103, 5338-5342
|Abstract »|Full Text »|PDF »
Dynamics of the Ras/ERK MAPK Cascade as Monitored by Fluorescent Probes.
A. Fujioka, K. Terai, R. E. Itoh, K. Aoki, T. Nakamura, S. Kuroda, E. Nishida, and M. Matsuda (2006)
J. Biol. Chem.
281, 8917-8926
|Abstract »|Full Text »|PDF »
Behavioral Profiling of Human Transitional Cell Carcinoma Ex vivo..
C. R. Estrada, M. Salanga, D. R. Bielenberg, W. B. Harrell, D. Zurakowski, X. Zhu, M. R. Palmer, M. R. Freeman, and R. M. Adam (2006)
Cancer Res.
66, 3078-3086
|Abstract »|Full Text »|PDF »
The Nuclear Import of Protein Kinase D3 Requires Its Catalytic Activity.
O. Rey, R. Papazyan, R. T. Waldron, S. H. Young, J. Lippincott-Schwartz, R. Jacamo, and E. Rozengurt (2006)
J. Biol. Chem.
281, 5149-5157
|Abstract »|Full Text »|PDF »
Microdomains of Intracellular Ca2+: Molecular Determinants and Functional Consequences.
Nuclear Trapping of the Forkhead Transcription Factor FoxO1 via Sirt-dependent Deacetylation Promotes Expression of Glucogenetic Genes.
D. Frescas, L. Valenti, and D. Accili (2005)
J. Biol. Chem.
280, 20589-20595
|Abstract »|Full Text »|PDF »
Fluorescence Imaging Reveals the Nuclear Behavior of Peroxisome Proliferator-activated Receptor/Retinoid X Receptor Heterodimers in the Absence and Presence of Ligand.
J. N. Feige, L. Gelman, C. Tudor, Y. Engelborghs, W. Wahli, and B. Desvergne (2005)
J. Biol. Chem.
280, 17880-17890
|Abstract »|Full Text »|PDF »
Applications of mRNA injections for analyzing cell lineage and asymmetric cell divisions during segmentation in the leech Helobdella robusta.
Analysis of baculovirus IE1 in living cells: dynamics and spatial relationships to viral structural proteins.
Y. Kawasaki, S. Matsumoto, and T. Nagamine (2004)
J. Gen. Virol.
85, 3575-3583
|Abstract »|Full Text »|PDF »
In Vivo Mammalian Brain Imaging Using One- and Two-Photon Fluorescence Microendoscopy.
J. C. Jung, A. D. Mehta, E. Aksay, R. Stepnoski, and M. J. Schnitzer (2004)
J Neurophysiol
92, 3121-3133
|Abstract »|Full Text »|PDF »
Insertion of Green Fluorescent Protein into Nonstructural Protein 5A Allows Direct Visualization of Functional Hepatitis C Virus Replication Complexes.
D. Moradpour, M. J. Evans, R. Gosert, Z. Yuan, H. E. Blum, S. P. Goff, B. D. Lindenbach, and C. M. Rice (2004)
J. Virol.
78, 7400-7409
|Abstract »|Full Text »|PDF »
Labeling of fusion proteins with synthetic fluorophores in live cells.
A. Keppler, H. Pick, C. Arrivoli, H. Vogel, and K. Johnsson (2004)
PNAS
101, 9955-9959
|Abstract »|Full Text »|PDF »
GFP-like Proteins as Ubiquitous Metazoan Superfamily: Evolution of Functional Features and Structural Complexity.
D. A. Shagin, E. V. Barsova, Y. G. Yanushevich, A. F. Fradkov, K. A. Lukyanov, Y. A. Labas, T. N. Semenova, J. A. Ugalde, A. Meyers, J. M. Nunez, et al. (2004)
Mol. Biol. Evol.
21, 841-850
|Abstract »|Full Text »|PDF »
Roles of the Proline-rich Domain in SLP-76 Subcellular Localization and T Cell Function.
A. L. Singer, S. C. Bunnell, A. E. Obstfeld, M. S. Jordan, J. N. Wu, P. S. Myung, L. E. Samelson, and G. A. Koretzky (2004)
J. Biol. Chem.
279, 15481-15490
|Abstract »|Full Text »|PDF »
From The Cover: Dynamics of HIV-1 recombination in its natural target cells.
D. N. Levy, G. M. Aldrovandi, O. Kutsch, and G. M. Shaw (2004)
PNAS
101, 4204-4209
|Abstract »|Full Text »|PDF »
Construction and characterization of a kappa opioid receptor devoid of all free cysteines.
D. Ott, R. Frischknecht, and A. Pluckthun (2004)
Protein Eng. Des. Sel.
17, 37-48
|Abstract »|Full Text »|PDF »
Session 8: Imaging/Tumor Dormancy: Imaging TONA M. GILMER, Department of Oncology Biology, GlaxoSmithKline, Research Triangle Park, North Carolina 27709.