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PMID: 2463257 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation of cell-to-cell adherens junctions from rat liver.

The Journal of cell biology ·Vol. 108 ·No. 1 ·1989-01-00 ·Pages 31-41

Tsukita S, Tsukita S

Abstract

A new isolation procedure for cell-to-cell adherens junctions has been developed using rat liver. From the bile canaliculi-enriched fraction obtained by homogenization of the liver and sucrose gradient centrifugation, the fraction rich in adherens junction was recovered by detergent treatment followed by sucrose gradient centrifugation. Light and electron microscopy revealed that this final fraction was mainly composed of the belt-like adherens junctions with their associated short actin filaments. Biochemical and immunological analyses have shown that vinculin is highly enriched in this fraction. Considering that vinculin is known to be localized in the cell-to-cell adherens junctions, we can conclude that we have succeeded in isolating the cell-to-cell adherens junctions. Furthermore, the constituents of the undercoat (dense layer underlying the membrane) of adherens junctions were selectively extracted from the fraction rich in junctions. Upon SDS electrophoresis of this extract, 10 polypeptides including vinculin, alpha-actinin, and actin were dominant. The results obtained are discussed with special reference to the molecular organization of the undercoats of cell-to-cell adherens junctions.

MeSH Terms
Actinin/analysis Actins/analysis Animals Bile Canaliculi/ultrastructure Bile Ducts, Intrahepatic/ultrastructure Cell Fractionation Centrifugation, Density Gradient Cytoskeletal Proteins Desmoplakins Desmosomes Intercellular Junctions/analysis,ultrastructure Keratins/analysis Liver/ultrastructure Membrane Glycoproteins/analysis Microscopy, Electron Muscle Proteins/analysis Rats Vinculin
Chemicals
Actins Cytoskeletal Proteins Desmoplakins Membrane Glycoproteins Muscle Proteins Actinin Vinculin Keratins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tsukita S
Department of Ultrastructural Research, Tokyo Metropolitan Institute of Medical Science, Japan.
Tsukita S
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39 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.108.1.31
Published
1989-01-00
Pages
31-41
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115366
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.108.1.31 PMC Full Text View journal details
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