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

被引:123
作者
Gaudry, CA
Verderio, E
Aeschlimann, D
Cox, A
Smith, C
Griffin, M
机构
[1] Nottingham Trent Univ, Dept Life Sci, Nottingham NG11 8NS, England
[2] Univ Wisconsin, Div Orthoped Surg, Madison, WI 53792 USA
[3] Unilever Res, Bedford, England
关键词
D O I
10.1074/jbc.274.43.30707
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 beta-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 beta-sandwich domain and that this interaction is crucial for cell surface association of tTG.
引用
收藏
页码:30707 / 30714
页数:8
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