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Specific fluorescent labeling of chicken myofibril Z-line proteins catalyzed by guinea pig liver transglutaminase

Gard, David L. and Lazarides, Elias (1979) Specific fluorescent labeling of chicken myofibril Z-line proteins catalyzed by guinea pig liver transglutaminase. Journal of Cell Biology, 81 (2). pp. 336-347. ISSN 0021-9525. http://resolver.caltech.edu/CaltechAUTHORS:GARjcb79

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Abstract

Guinea pig liver transglutaminase has been found to catalyze the covalent incorporation of dansylcadaverine into chicken skeletal muscle myofibril proteins. Epifluorescence microscopy reveals that the incorporated dansylcadaverine is specifically localized at or near the myofibril Z line. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) indicates that actin constitutes a major fraction of the labeled material; the Z-line proteins alpha-actinin and desmin also show significant labeling, as well as tropomyosin, several additional unidentified proteins, and material with an extremely high molecular weight. The Z-line-specific fluorescence can be removed by brief trypsinization, which releases fluorescent alpha-actinin into the supernate. The majority of the fluorescent protein species are resistant to extraction by either 0.6 M KCl or KI. These results, in conjunction with the microscopic localization, suggest that the dansyl- labeled proteins are constituents of the myofibril Z line. A significant amount of fluorescently labeled transglutaminase is also present in labeled myofibrils, which is resistant to extraction with either 0.6 M KCl or KI. This result indicates a strong, noncovalent interaction between the transglutaminase molecule and the myofibril Z line.


Item Type:Article
Related URLs:
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http://www.jcb.org/cgi/content/abstract/81/2/336PublisherUNSPECIFIED
Additional Information:Copyright © 1979 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. Received for publication 17 April 1978, and in revised form 18 December 1978. We thank Mr. David R. Balzer, Jr., for his assistance with two-dimensional gel electrophoresis. 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. David L. Gard was also supported by a National Science Foundation Predoctoral Fellowship.
Funders:
Funding AgencyGrant Number
National Institutes of HealthPHS GM 06965-18
Muscular Dystrophy Association of AmericaUNSPECIFIED
American Cancer SocietyUNSPECIFIED
National Science Foundation predoctoral fellowshipUNSPECIFIED
Subject Keywords:actin; desmin; α-actinin
Record Number:CaltechAUTHORS:GARjcb79
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:GARjcb79
Usage Policy:RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.
ID Code:11381
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:11 Aug 2008 18:44
Last Modified:26 Dec 2012 10:13

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