Intimin-mediated export of passenger proteins requires maintenance of a translocation-competent conformation

被引:26
作者
Adams, TM [1 ]
Wentzel, A [1 ]
Kolmar, H [1 ]
机构
[1] Univ Gottingen, Abt Mol Genet & Praparat Mol Biol, Inst Mikrobiol & Genet, D-37077 Gottingen, Germany
关键词
D O I
10.1128/JB.187.2.522-533.2005
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Intimins from pathogenic bacteria promote intimate bacterial adhesion to epithelial cells. Several structurally similar domains form on the bacterial cell surface an extended rigid rod that exposes the carboxy-terminal domain, which interacts with the translocated intimin receptor. We constructed a series of intimin-derived fusion proteins consisting of carboxy-terminally truncated intimin and the immunoglobulin light-chain variable domain REIv, ubiquitin, calmodulin, beta-lactamase inhibitor protein, or betan-lactamase. By systematically investigating the intimin-mediated cell surface exposure of these passenger domains in the presence or absence of compounds that interfere with outer membrane stability or passenger domain folding, we acquired experimental evidence that intimin-mediated protein export across the outer membrane requires, prior to export, the maintenance of a translocation-competent conformation that may be distinct from the final protein structure. We propose that, during export, competition exists between productive translocation and folding of the passenger domain in the periplasm into a stable conformation that is not compatible with translocation through the bacterial outer membrane. These results may expand understanding of the mechanism by which intimins are inserted into the outer membrane and expose extracellular domains on the cell surface.
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收藏
页码:522 / 533
页数:12
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[1]   Detection of intimins α, β, γ, and δ, four intimin derivatives expressed by attaching and effacing microbial pathogens [J].
Adu-Bobie, J ;
Frankel, G ;
Bain, C ;
Goncalves, AG ;
Trabulsi, LR ;
Douce, G ;
Knutton, S ;
Dougan, G .
JOURNAL OF CLINICAL MICROBIOLOGY, 1998, 36 (03) :662-668
[2]  
AKIYAMA Y, 1992, J BIOL CHEM, V267, P22440
[3]   IDENTIFICATION OF A PROTEIN REQUIRED FOR DISULFIDE BOND FORMATION INVIVO [J].
BARDWELL, JCA ;
MCGOVERN, K ;
BECKWITH, J .
CELL, 1991, 67 (03) :581-589
[4]   Structural basis for recognition of the translocated intimin receptor (Tir) by intimin from enteropathogenic Escherichia coli [J].
Batchelor, M ;
Prasannan, S ;
Daniell, S ;
Reece, S ;
Connerton, I ;
Bloomberg, G ;
Dougan, G ;
Frankel, G ;
Matthews, S .
EMBO JOURNAL, 2000, 19 (11) :2452-2464
[5]   CLONING AND EXPRESSION OF AN ADHESIN (AIDA-I) INVOLVED IN DIFFUSE ADHERENCE OF ENTEROPATHOGENIC ESCHERICHIA-COLI [J].
BENZ, I ;
SCHMIDT, MA .
INFECTION AND IMMUNITY, 1989, 57 (05) :1506-1511
[6]   Periplasmic transit and disulfide bond formation of the autotransported Shigella protein IcsA [J].
Brandon, LD ;
Goldberg, MB .
JOURNAL OF BACTERIOLOGY, 2001, 183 (03) :951-958
[7]   The general protein secretory pathway: phylogenetic analyses leading to evolutionary conclusions [J].
Cao, TB ;
Saier, MH .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 2003, 1609 (01) :115-125
[8]   Type IV secretion: intercellular transfer of macromolecules by systems ancestrally related to conjugation machines [J].
Christie, PJ .
MOLECULAR MICROBIOLOGY, 2001, 40 (02) :294-305
[9]   The cystine knot of a squash-type protease inhibitor as a structural scaffold for Escherichia coli cell surface display of conformationally constrained peptides [J].
Christmann, A ;
Walter, K ;
Wentzel, A ;
Krätzner, R ;
Kolmar, H .
PROTEIN ENGINEERING, 1999, 12 (09) :797-806
[10]   The vacuolating cytotoxin of Helicobacter pylori [J].
Cover, TL .
MOLECULAR MICROBIOLOGY, 1996, 20 (02) :241-246