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Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments

Hubbard, Bruce D. and Lazarides, Elias (1979) Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments. Journal of Cell Biology, 80 (1). pp. 166-182. ISSN 0021-9525. http://resolver.caltech.edu/CaltechAUTHORS:HUBjcb79

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Abstract

Desmin is a 50,000-mol wt protein that is enriched along with 100-A filaments in chicken gizzard that has been extracted with 1 M KI. Although 1 M KI removes most of the actin from gizzard, a small fraction of this protein remains persistently insoluble, along with desmin. The solubility properties of this actin are the same as for desmin: they are both insoluble in high salt concentrations, but are solubilized at low pH or by agents that dissociate hydrophobic bonds. Desmin may be purified by repeated cycles of solubilization by 1 M acetic acid and subsequent precipitation by neutralization to pH 4. During this process, a constant nonstoichiometric ratio of actin to desmin is attained. Gel filtration on Ultrogel AcA34 in the presence of 0.5% Sarkosyl NL-97 reveals nonmonomeric fractions of actin and desmin that comigrate through the column. Gel filtration on Bio-Gel P300 in the presence of 1 M acetic acid reveals that the majority of desmin is monomeric under these conditions. A small fraction of desmin and all of the actin elute with the excluded volume. When the acetic acid is removed from actin-desmin solutions by dialysis, a gel forms that is composed of filaments with diameters of 120-140 A. These filaments react uniformly with both anti-actin and anti-desmin antiserum. These results suggest that desmin is the major subunit of the muscle 100-A filaments and that it may form nonstoichiometric complexes with actin.


Item Type:Article
Additional Information:Copyright © 1979 by The Rockefeller University Press. Received for publication 16 March 1978, and in revised form 11 July 1978. We would like to thank the two reviewers for their time and effort and their many helpful comments. This work was supported by grant PHS-GM 06965-18 from the National Institutes of Health, and also by grants from the Muscular Dystrophy Association of America and the American Cancer Society.
Subject Keywords:100 A filaments;immunofluorescence; α-actinin; Z disk
Record Number:CaltechAUTHORS:HUBjcb79
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:HUBjcb79
Alternative URL:http://www.jcb.org/cgi/content/abstract/80/1/166
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:4977
Collection:CaltechAUTHORS
Deposited By: Lindsay Cleary
Deposited On:18 Sep 2006
Last Modified:26 Dec 2012 09:02

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