Cell surface localization of tissue transglutaminase is dependent on a fibronectin-binding site in its N-terminal beta-sandwich domain

Gaudry, CA, Verderio, E ORCID: 0000-0001-9153-8997, Aeschlimann, D, Cox, A, Smith, C and Griffin, M, 1999. Cell surface localization of tissue transglutaminase is dependent on a fibronectin-binding site in its N-terminal beta-sandwich domain. Journal of Biological Chemistry, 274 (43), pp. 30707-30714. ISSN 0021-9258

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Abstract

Increasing evidence indicates that tissue transglutaminase (tTG) plays a role in the assembly and remodeling of extracellular matrices and promotes cell adhesion. Using an inducible system we have previously shown that tTG associates with the extracellular matrix deposited by stably transfected 3T3 fibroblasts overexpressing the enzyme. We now show by confocal microscopy that tTG colocalizes with pericellular fibronectin in these cells, and by immunogold electron microscopy that the two proteins are found in clusters at the cell surface. Expression vectors encoding the full-length tTG or a N-terminal truncated tTG lacking the proposed fibronectin-binding site (fused to the bacterial reporter enzyme β-galactosidase) were generated to characterize the role of fibronectin in sequestration of tTG in the pericellular matrix. Enzyme-linked immunosorbent assay style procedures using extracts of transiently transfected COS-7 cells and immobilized fibronectin showed that the truncation abolished fibronectin binding. Similarly, the association of tTG with the pericellular matrix of cells in suspension or with the extracellular matrix deposited by cell monolayers was prevented by the truncation. These results demonstrate that tTG binds to the pericellular fibronectin coat of cells via its N-terminal β-sandwich domain and that this interaction is crucial for cell surface association of tTG.

Item Type: Journal article
Publication Title: Journal of Biological Chemistry
Creators: Gaudry, C.A., Verderio, E., Aeschlimann, D., Cox, A., Smith, C. and Griffin, M.
Publisher: The American Society for Biochemistry and Molecular Biology, Inc.
Date: 1999
Volume: 274
Number: 43
ISSN: 0021-9258
Identifiers:
NumberType
10.1074/jbc.274.43.30707DOI
Rights: Copyright © 1999 by American Society for Biochemistry and Molecular Biology, Inc.
Divisions: Schools > School of Science and Technology
Depositing User: EPrints Services
Date Added: 09 Oct 2015 09:40
Last Modified: 09 Jun 2017 13:07
URI: http://irep.ntu.ac.uk/id/eprint/807

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