A crucial sequence for transglutaminase type 2 extracellular trafficking in renal tubular epithelial cells lies in its N-terminal {beta}-sandwich domain

Tools

Chou, CY, Streets, AJ, Watson, PF, Huang, L, Verderio, E ORCID: https://orcid.org/0000-0001-9153-8997 and Johnson, TS, 2011. A crucial sequence for transglutaminase type 2 extracellular trafficking in renal tubular epithelial cells lies in its N-terminal {beta}-sandwich domain. Journal of Biological Chemistry, 286, pp. 27825-27835.

[thumbnail of 205524_7680 Verderio-Edwards Publisher.pdf]

Preview

Text
205524_7680 Verderio-Edwards Publisher.pdf
Download (3MB) | Preview

Abstract

Transglutaminase type 2 (TG2) catalyzes the formation of an -( -glutamyl)-lysine isopeptide bond between adjacent peptides or proteins including those of the extracellular matrix (ECM). Elevated extracellular TG2 leads to accelerated ECM deposition and reduced clearance that underlie tissue scarring and fibrosis. The extracellular trafficking of TG2 is crucial to its role in ECM homeostasis; however, the mechanism by which TG2 escapes the cell is unknown as it has no signal leader peptide and therefore cannot be transported classically. Understanding TG2 transport may highlight novel mechanisms to interfere with the extracellular function of TG2 as isoform-specific TG2 inhibitors remain elusive. Mammalian expression vectors were constructed containing domain deletions of TG2. These were transfected into three kidney tubular epithelial cell lines, and TG2 export was assessed to identify critical domains. Point mutation was then used to highlight specific sequences within the domain required for TG2 export. The removal of -sandwich domain prevented all TG2 export. Mutations of Asp94 and Asp97 within the N-terminal -sandwich domain were identified as crucial for TG2 externalization. These form part of a previously identified fibronectin binding domain (88WTATVVDQQDCTLSLQLTT106). However, siRNA knockdown of fibronectin failed to affect TG2 export. The sequence 88WTATVVDQQDCTLSLQLTT106 within the -sandwich domain of TG2 is critical to its export in tubular epithelial cell lines. The extracellular trafficking of TG2 is independent of fibronectin.

Item Type:	Journal article
Publication Title:	Journal of Biological Chemistry
Creators:	Chou, C.Y., Streets, A.J., Watson, P.F., Huang, L., Verderio, E. and Johnson, T.S.
Publisher:	American Society for Biochemistry and Molecular Biology
Date:	2011
Volume:	286
Rights:	© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.
Divisions:	Schools > School of Science and Technology
Record created by:	EPrints Services
Date Added:	09 Oct 2015 11:07
Last Modified:	09 Jun 2017 13:50
URI:	https://irep.ntu.ac.uk/id/eprint/23150