Cross-linking of Novikoff ascites hepatoma cytokeratin filaments

W. S WARD; W. N SCHMIDT; C. A SCHMIDT; L. S HNILICA

doi:10.1021/bi00337a026

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Cross-linking of Novikoff ascites hepatoma cytokeratin filaments

Journal article

Peer reviewed

Cross-linking of Novikoff ascites hepatoma cytokeratin filaments

W. S WARD, W. N SCHMIDT, C. A SCHMIDT and L. S HNILICA

Biochemistry (Easton), Vol.24(16), pp.4429-4434

1985

DOI: 10.1021/bi00337a026

PMID: 2413887

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Abstract

We have investigated the structure of solubilized cytokeratins from Novikoff ascites hepatoma using the cleavable cross-linker 3,3'-dithiobis(sulfosuccinimidyl propionate) in the presence of 6 M urea to effect partial complex melting. By two-dimensional gel electrophoresis, in which the protein cross-links were broken in the second dimension, we have identified two major complexes as a p39-p56 dimer and a (p39-p56)2 tetramer, p39 and p56 being two of the major cytokeratins in Novikoff ascites hepatoma. Experiments investigating possible relationships between the dimer and tetramer employed immunoblots and two monoclonal antibodies which recognized either p56 or p39 cytokeratins. When very low protein concentrations were cross-linked, the dimer was the predominant product. As protein concentration increased, we noted a decrease in dimers and a corresponding increase in tetramers, suggesting that the dimer may be a precursor to the tetramer. In support of the cross-linking experiments, two-dimensional gel electrophoresis using 4 M urea in the first dimension indicated a predominant association of p56 and p39 in the Novikoff ascites hepatoma cytokeratin complexes.

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Cytoskeleton, cytoplasm. Intracellular movements

Molecular and cellular biology

Cell structures and functions

Details

Title: Subtitle: Cross-linking of Novikoff ascites hepatoma cytokeratin filaments
Creators: W. S WARD - Vanderbilt univ., school medicine
W. N SCHMIDT - Vanderbilt univ., school medicine
C. A SCHMIDT - Vanderbilt univ., school medicine
L. S HNILICA - Vanderbilt univ., school medicine
Resource Type: Journal article
Publication Details: Biochemistry (Easton), Vol.24(16), pp.4429-4434
Publisher: American Chemical Society
DOI: 10.1021/bi00337a026
PMID: 2413887
ISSN: 0006-2960
eISSN: 1520-4995
Language: English
Date published: 1985
Academic Unit: Gastroenterology and Hepatology; Internal Medicine
Record Identifier: 9984094314102771

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