Secretion of the intimin passenger domain is driven by protein folding

Leo, J.C. ORCID: 0000-0002-7066-7527, Oberhettinger, P., Yoshimoto, S., Udatha, D.B.R.K.G., Morth, J.P., Schütz, M., Hori, K. and Linke, D., 2016. Secretion of the intimin passenger domain is driven by protein folding. Journal of Biological Chemistry, 291 (38), pp. 20096-20112. ISSN 1083-351X

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Abstract

Intimin is an essential adhesin of attaching and effacing organisms such as entropathogenic Escherichia coli. It is also the prototype of type Ve secretion or inverse autotransport, where the extracellular C-terminal region or passenger is exported with the help of an N-terminal transmembrane -barrel domain. We recently reported a stalled secretion intermediate of intimin, where the passenger is located in the periplasm but the -barrel is already inserted into the membrane. Stalling of this mutant is due to the insertion of an epitope tag at the very N terminus of the passenger. Here, we examined how this insertion disrupts autotransport and found that it causes misfolding of the N-terminal immunoglobulin (Ig)-like domain D00. We could also stall the secretion by making an internal deletion in D00, and introducing the epitope tag into the second Ig-like domain, D0, also resulted in reduced passenger secretion. In contrast to many classical autotransporters, where a proximal folding core in the passenger is required for secretion, the D00 domain is dispensable, as the passenger of an intimin mutant lacking D00 entirely is efficiently exported. Furthermore, the D00 domain is slightly less stable than the D0 and D1 domains, unfolding at 200 piconewtons (pN) compared with 250 pN for D0 and D1 domains as measured by atomic force microscopy. Our results support a model where the secretion of the passenger is driven by sequential folding of the extracellular Ig-like domains, leading to vectorial transport of the passenger domain across the outer membrane in an N to C direction.

Item Type: Journal article
Publication Title: Journal of Biological Chemistry
Creators: Leo, J.C., Oberhettinger, P., Yoshimoto, S., Udatha, D.B.R.K.G., Morth, J.P., Schütz, M., Hori, K. and Linke, D.
Publisher: American Society for Biochemistry and Molecular Biology (ASBMB)
Date: 16 September 2016
Volume: 291
Number: 38
ISSN: 1083-351X
Identifiers:
NumberType
10.1074/jbc.m116.731497DOI
Rights: © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
Divisions: Schools > School of Science and Technology
Depositing User: Jill Tomkinson
Date Added: 18 Jun 2019 12:45
Last Modified: 25 Jun 2019 13:04
URI: http://irep.ntu.ac.uk/id/eprint/36840

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