QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online June 14, 2004
doi: 10.1242/10.1242/jcs.01167

This Article Figures Only Full Text Full Text (PDF) Supplemental Data Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Peet, J. A. Articles by Pugh, E. N. Search for Related Content PubMed PubMed Citation Articles by Peet, J. A. Articles by Pugh, E. N., Jr Social Bookmarking                         What's this?

Quantification of the cytoplasmic spaces of living cells with EGFP reveals arrestin-EGFP to be in disequilibrium in dark adapted rod photoreceptors

¹ Temple University School of Medicine, 3400 North Broad Street, Philadelphia, PA 19140, USA
² F. M. Kirby Center for Molecular Ophthalmology, University of Pennsylvania School of Medicine, Stellar-Chance Building, Room 309B, 422 Curie Boulevard, PA 19104-6069, USA
³ Department of Biochemistry and Molecular Biology, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, NY 13210-2375, USA
⁴ Department of Ophthalmology, SUNY Upstate Medical University, 750 East Adams Street, Syracuse, NY 13210-2375, USA
⁵ Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226-0509, USA

^* Author for correspondence (e-mail: pugh{at}mail.med.upenn.edu)

Accepted 19 February 2004

The hypothesis is tested that enhanced green fluorescent protein (EGFP) can be used to quantify the aqueous spaces of living cells, using as a model transgenic Xenopus rods. Consistent with the hypothesis, regions of rods having structures that exclude EGFP, such as the mitochondrial-rich ellipsoid and the outer segments, have highly reduced EGFP fluorescence. Over a 300-fold range of expression the average EGFP concentration in the outer segment was approximately half that in the most intensely fluorescent regions of the inner segment, in quantitative agreement with prior X-ray diffraction estimates of outer segment cytoplasmic volume. In contrast, the fluorescence of soluble arrestin-EGFP fusion protein in the dark adapted rod outer segment was approximately threefold lower than predicted by the EGFP distribution, establishing that the fusion protein is not equilibrated with the cytoplasm. Arrestin-EGFP mass was conserved during a large-scale, light-driven redistribution in which 40% of the protein in the inner segment moved to the outer segment in less than 30 minutes.

Key words: Protein movement, Arrestin, Xenopus, Transgenesis

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				S. A. Baker, M. Haeri, P. Yoo, S. M. Gospe III, N. P. Skiba, B. E. Knox, and V. Y. Arshavsky The outer segment serves as a default destination for the trafficking of membrane proteins in photoreceptors J. Cell Biol., November 3, 2008; 183(3): 485 - 498. [Abstract] [Full Text] [PDF]

				K. Kam and R. Nicoll Excitatory Synaptic Transmission Persists Independently of the Glutamate Glutamine Cycle J. Neurosci., August 22, 2007; 27(34): 9192 - 9200. [Abstract] [Full Text] [PDF]

				S. Nickell, P. S.-H. Park, W. Baumeister, and K. Palczewski Three-dimensional architecture of murine rod outer segments determined by cryoelectron tomography J. Cell Biol., June 21, 2007; 177(5): 917 - 925. [Abstract] [Full Text] [PDF]

				D. H. Rosenzweig, K. S. Nair, J. Wei, Q. Wang, G. Garwin, J. C. Saari, C.-K. Chen, A. V. Smrcka, A. Swaroop, J. Lem, et al. Subunit Dissociation and Diffusion Determine the Subcellular Localization of Rod and Cone Transducins J. Neurosci., May 16, 2007; 27(20): 5484 - 5494. [Abstract] [Full Text] [PDF]

				S. E. Haire, J. Pang, S. L. Boye, I. Sokal, C. M. Craft, K. Palczewski, W. W. Hauswirth, and S. L. Semple-Rowland Light-Driven Cone Arrestin Translocation in Cones of Postnatal Guanylate Cyclase-1 Knockout Mouse Retina Treated with AAV-GC1. Invest. Ophthalmol. Vis. Sci., September 1, 2006; 47(9): 3745 - 3753. [Abstract] [Full Text] [PDF]

				K. J. Strissel, M. Sokolov, L. H. Trieu, and V. Y. Arshavsky Arrestin Translocation Is Induced at a Critical Threshold of Visual Signaling and Is Superstoichiometric to Bleached Rhodopsin J. Neurosci., January 25, 2006; 26(4): 1146 - 1153. [Abstract] [Full Text] [PDF]

				J. J. Peterson, W. Orisme, J. Fellows, J. H. McDowell, C. L. Shelamer, D. R. Dugger, and W. C. Smith A Role for Cytoskeletal Elements in the Light-Driven Translocation of Proteins in Rod Photoreceptors Invest. Ophthalmol. Vis. Sci., November 1, 2005; 46(11): 3988 - 3998. [Abstract] [Full Text] [PDF]