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A biochemical dissection of the cardiac intercalated disk: isolation of subcellular fractions containing fascia adherentes and gap junctions

Colaco, C. A. and Evans, William Howard 1981. A biochemical dissection of the cardiac intercalated disk: isolation of subcellular fractions containing fascia adherentes and gap junctions. Journal of Cell Science 52 , pp. 313-325.

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Abstract

In view of our limited knowledge of the biochemical composition of intercellular junctions, a method was developed for the preparation from rats and mice of plasma membranes containing cardiac intercalated disks. When these membranes were extracted with detergents, e.g. N-lauryl sarcosinate or deoxycholate, the detergent-insoluble material contained structures derived mainly from fascia adherentes junctions, but a few gap junctions and maculae adherentes were also present. When the detergent extraction was carried out at an alkaline pH, the maculae adherentes junctions were dissolved. Fractionation of the detergent-insoluble extract on a sucrose gradient yielded a fraction containing fascia adherentes junction of density 1.20-1.26 g/cm3. Gap junctions banded at a lower density, 1.16-1.20 g/cm3. Polyacrylamide gel electrophoresis showed that the major polypeptide bands in the fascia adherentes-enriched fraction were of molecular weights 134000, 108000, 62-64000, 58000, 47000 and 43000. Although fractions with the gap junctions were contaminated by fascia adherentes junctions, the major polypeptides were calculated by subtraction to be of mol. wt 37000, 26000 and 19000. Two glycoproteins corresponding to minor polypeptides visualized by Coomassie Blue staining were present in the fascia adherentes fraction. Comparison of the fascia adherentes-enriched fraction with a Z-disc fraction prepared from rabbit hearts indicated a different morphology and polypeptide composition.

Item Type: Article
Status: Published
Schools: Medicine
Subjects: R Medicine > R Medicine (General)
R Medicine > RZ Other systems of medicine
Uncontrolled Keywords: Animals,Detergents,ElectrophoresisPolyacrylamideGel,IntercellularJunctions/analysis*,IntercellularJunctions/ultrastructure,Mice,MicroscopyElectron,Myocardium/analysis*,Myocardium/ultrastructure,Peptides/analysis,Rabbits,Rats,Sarcolemma/analysis Substances Detergents,Peptides
Additional Information: Publication Types Research Support, Non-U.S. Gov't Full Text Sources HighWire - PDF Other Literature Sources Access more work from the authors - ResearchGate
Publisher: The Company of Biologists Ltd
ISSN: 0021-9533
Last Modified: 17 Mar 2021 02:53
URI: https://orca.cardiff.ac.uk/id/eprint/66156

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