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Studying protein dynamics in living cells.
Lippincott-Schwartz J, Snapp E, Kenworthy A. Lippincott-Schwartz J, et al. Nat Rev Mol Cell Biol. 2001 Jun;2(6):444-56. doi: 10.1038/35073068. Nat Rev Mol Cell Biol. 2001. PMID: 11389468 Review.
Dynamics of putative raft-associated proteins at the cell surface.
Kenworthy AK, Nichols BJ, Remmert CL, Hendrix GM, Kumar M, Zimmerberg J, Lippincott-Schwartz J. Kenworthy AK, et al. J Cell Biol. 2004 Jun 7;165(5):735-46. doi: 10.1083/jcb.200312170. Epub 2004 Jun 1. J Cell Biol. 2004. PMID: 15173190 Free PMC article.
Here, we use fluorescence recovery after photobleaching to test if raft association affects a protein's ability to laterally diffuse large distances across the cell surface. ...
Here, we use fluorescence recovery after photobleaching to test if raft association affects a protein's ability to laterally diffuse …
Depalmitoylated Ras traffics to and from the Golgi complex via a nonvesicular pathway.
Goodwin JS, Drake KR, Rogers C, Wright L, Lippincott-Schwartz J, Philips MR, Kenworthy AK. Goodwin JS, et al. J Cell Biol. 2005 Jul 18;170(2):261-72. doi: 10.1083/jcb.200502063. J Cell Biol. 2005. PMID: 16027222 Free PMC article.
A combination of time-lapse microscopy and photobleaching techniques reveal that in the absence of palmitoylation, GFP-tagged HRas and NRas undergo rapid exchange between the cytosol and ER/Golgi membranes, and that wild-type GFP-HRas and GFP-NRas are recycled to the Golgi
A combination of time-lapse microscopy and photobleaching techniques reveal that in the absence of palmitoylation, GFP-tagged HRas an
Analysis of diffusion in curved surfaces and its application to tubular membranes.
Klaus CJ, Raghunathan K, DiBenedetto E, Kenworthy AK. Klaus CJ, et al. Mol Biol Cell. 2016 Dec 1;27(24):3937-3946. doi: 10.1091/mbc.E16-06-0445. Epub 2016 Oct 12. Mol Biol Cell. 2016. PMID: 27733625 Free PMC article.
Diffusion of particles in curved surfaces is inherently complex compared with diffusion in a flat membrane, owing to the nonplanarity of the surface. ...In addition, we show that tubular structures tend to retain concentration gradients for a longer time compared wi …
Diffusion of particles in curved surfaces is inherently complex compared with diffusion in a flat membrane, owing to the nonplanarity …
Myosin Vb interacts with Rab8a on a tubular network containing EHD1 and EHD3.
Roland JT, Kenworthy AK, Peranen J, Caplan S, Goldenring JR. Roland JT, et al. Mol Biol Cell. 2007 Aug;18(8):2828-37. doi: 10.1091/mbc.e07-02-0169. Epub 2007 May 16. Mol Biol Cell. 2007. PMID: 17507647 Free PMC article.
Rab8a and Myosin Vb colocalize to a tubular network containing EHD1 and EHD3, which does not contain Rab11a. Myosin Vb tail can cause the accumulation of both Rab11a and Rab8a in collapsed membrane cisternae, whereas dominant-negative Rab11-FIP2(129-512) selectively accumu …
Rab8a and Myosin Vb colocalize to a tubular network containing EHD1 and EHD3, which does not contain Rab11a. Myosin Vb tail can cause …
Photobleaching approaches to investigate diffusional mobility and trafficking of Ras in living cells.
Goodwin JS, Kenworthy AK. Goodwin JS, et al. Methods. 2005 Oct;37(2):154-64. doi: 10.1016/j.ymeth.2005.05.013. Methods. 2005. PMID: 16288889
While all of these techniques exploit photobleaching as a tool to monitor protein dynamics, they each provide a unique subset of information. In particular, FRAP provides measurements of protein mobility via lateral diffusion by monitoring recovery of fluorescence i …
While all of these techniques exploit photobleaching as a tool to monitor protein dynamics, they each provide a unique subset …
Analysis of protein and lipid dynamics using confocal fluorescence recovery after photobleaching (FRAP).
Day CA, Kraft LJ, Kang M, Kenworthy AK. Day CA, et al. Curr Protoc Cytom. 2012 Oct;Chapter 2:Unit2.19. doi: 10.1002/0471142956.cy0219s62. Curr Protoc Cytom. 2012. PMID: 23042527 Free PMC article.
In this unit, we discuss the theoretical basis for confocal FRAP, followed by step-by-step protocols for FRAP data acquisition using a laser-scanning confocal microscope for (1) measuring the diffusion of a membrane protein, (2) measuring the diffusion of a s …
In this unit, we discuss the theoretical basis for confocal FRAP, followed by step-by-step protocols for FRAP data acquisition using a
Imaging protein complex formation in the autophagy pathway: analysis of the interaction of LC3 and Atg4B(C74A) in live cells using Förster resonance energy transfer and fluorescence recovery after photobleaching.
Kraft LJ, Kenworthy AK. Kraft LJ, et al. J Biomed Opt. 2012 Jan;17(1):011008. doi: 10.1117/1.JBO.17.1.011008. J Biomed Opt. 2012. PMID: 22352642 Free PMC article.
The protein microtubule-associated protein 1, light chain 3 (LC3) functions in autophagosome formation and plays a central role in the autophagy pathway. Previously, we found LC3 diffuses more slowly in cells than is expected for a freely diffusing monomer, suggesti …
The protein microtubule-associated protein 1, light chain 3 (LC3) functions in autophagosome formation and plays a central role in th …
The lateral mobility of NHE3 on the apical membrane of renal epithelial OK cells is limited by the PDZ domain proteins NHERF1/2, but is dependent on an intact actin cytoskeleton as determined by FRAP.
Cha B, Kenworthy A, Murtazina R, Donowitz M. Cha B, et al. J Cell Sci. 2004 Jul 1;117(Pt 15):3353-65. doi: 10.1242/jcs.01180. J Cell Sci. 2004. PMID: 15226406
The epithelial brush border (BB) Na+/H+ exchanger, NHE3, plays a major role in transcellular Na+ absorption in the renal proximal tubule. ...A confocal microscope was used to allow the selective study of apical membrane versus intracellular NHE3. A chimera of …
The epithelial brush border (BB) Na+/H+ exchanger, NHE3, plays a major role in transcellular Na+ absorption in the renal proximal tub …
Quantitative electron microscopy and fluorescence spectroscopy of the membrane distribution of influenza hemagglutinin.
Hess ST, Kumar M, Verma A, Farrington J, Kenworthy A, Zimmerberg J. Hess ST, et al. J Cell Biol. 2005 Jun 20;169(6):965-76. doi: 10.1083/jcb.200412058. J Cell Biol. 2005. PMID: 15967815 Free PMC article.
Here, we use influenza viral envelope protein hemagglutinin (HA(0)) to test the hypothesis that clustering results from proteins partitioning into preexisting, fluid-ordered "raft" domains, wherein they have a random distribution. ...The magnitude and density dependence of …
Here, we use influenza viral envelope protein hemagglutinin (HA(0)) to test the hypothesis that clustering results from proteins partitionin …
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