Multicolor and Electron Microscopic Imaging of Connexin Trafficking

Recombinant proteins containing tetracysteine tags can be successively labeled in living cells with different colors of biarsenical fluorophores so that older and younger protein molecules can be sharply distinguished by both fluorescence and electron microscopy. Here we used this approach to show t...

Full description

Saved in:
Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2002-04, Vol.296 (5567), p.503-507
Main Authors: Gaietta, Guido, Deerinck, Thomas J., Adams, Stephen R., Bouwer, James, Tour, Oded, Laird, Dale W., Sosinsky, Gina E., Tsien, Roger Y., Ellisman, Mark H.
Format: Article
Language:English
Subjects:
3,3'-Diaminobenzidine - chemistry
Amino Acid Motifs
Animals
Arsenicals - metabolism
Biological and medical sciences
Cell Line
Cell Membrane - metabolism
Cell Membrane - ultrastructure
Cell membranes
Cell structures and functions
Cells
Connexin 43 - biosynthesis
Connexin 43 - metabolism
Connexins
Cysteine - chemistry
Electron microscopes
Electron microscopy
Electrons
Endocytosis
Exocytosis
Fluoresceins - metabolism
Fluorescence
Fluorescent Dyes - metabolism
Fundamental and applied biological sciences. Psychology
Gap junctions
Gap Junctions - metabolism
Gap Junctions - ultrastructure
HeLa Cells
Humans
Imaging
Literary Devices
Microscopes
Microscopy, Confocal
Microscopy, Electron
Microscopy, Immunoelectron
Miscellaneous
Molecular and cellular biology
Molecules
Organometallic Compounds - metabolism
Oxazines - metabolism
Patch-Clamp Techniques
Protein Transport
Proteins
Recombinant proteins
Recombinant Proteins - metabolism
Transport Vesicles - metabolism
Transport Vesicles - ultrastructure
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Recombinant proteins containing tetracysteine tags can be successively labeled in living cells with different colors of biarsenical fluorophores so that older and younger protein molecules can be sharply distinguished by both fluorescence and electron microscopy. Here we used this approach to show that newly synthesized connexin43 was transported predominantly in 100- to 150-nanometer vesicles to the plasma membrane and incorporated at the periphery of existing gap junctions, whereas older connexins were removed from the center of the plaques into pleiomorphic vesicles of widely varying sizes. Selective imaging by correlated optical and electron microscopy of protein molecules of known ages will clarify fundamental processes of protein trafficking in situ.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.1068793