Size and conformation limits to secretion of disulfide-bonded loops in autotransporter proteins.

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@article{a7966e861d1240739719f3642a9c737a,
title = "Size and conformation limits to secretion of disulfide-bonded loops in autotransporter proteins.",
abstract = "Autotransporters are a super-family of virulence factors typified by a channel-forming C-terminus that facilitates translocation of the functional N-terminal passenger domain across the outer membrane of Gram-negative bacteria. This final step in the secretion of autotransporters requires a translocation-competent conformation for the passenger domain that differs markedly from the structure of the fully folded secreted protein. The nature of the translocation-competent conformation remains controversial, in particular whether the passenger domain can adopt secondary structural motifs, such as disulfide-bonded segments, while maintaining a secretion competent state. Here we use the endogenous and closely spaced cysteine residues of the Pet toxin from enteroaggregative E. coli to investigate the effect of disulfide-bond-induced folding on translocation of an autotransporter passenger domain. We reveal that rigid structural elements within disulfide-bonded segments are resistant to autotransporter-mediated secretion. We define the size limit of disulfide-bonded segments tolerated by the autotransporter system demonstrating that, when present, cysteine pairs are intrinsically closely spaced to prevent congestion of the translocator pore by large disulfide-bonded regions. These latter data strongly support the hairpin mode of autotransporter biogenesis.",
author = "Denisse Leyton and Yanina Sevastsyanovich and Douglas Browning and Amanda Rossiter and Timothy Wells and RE Fitzpatrick and Michael Overduin and Adam Cunningham and Ian Henderson",
year = "2011",
month = oct
day = "17",
doi = "10.1074/jbc.M111.306118",
language = "English",
volume = "286",
pages = "42283--42291",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "49",

}

RIS

TY - JOUR

T1 - Size and conformation limits to secretion of disulfide-bonded loops in autotransporter proteins.

AU - Leyton, Denisse

AU - Sevastsyanovich, Yanina

AU - Browning, Douglas

AU - Rossiter, Amanda

AU - Wells, Timothy

AU - Fitzpatrick, RE

AU - Overduin, Michael

AU - Cunningham, Adam

AU - Henderson, Ian

PY - 2011/10/17

Y1 - 2011/10/17

N2 - Autotransporters are a super-family of virulence factors typified by a channel-forming C-terminus that facilitates translocation of the functional N-terminal passenger domain across the outer membrane of Gram-negative bacteria. This final step in the secretion of autotransporters requires a translocation-competent conformation for the passenger domain that differs markedly from the structure of the fully folded secreted protein. The nature of the translocation-competent conformation remains controversial, in particular whether the passenger domain can adopt secondary structural motifs, such as disulfide-bonded segments, while maintaining a secretion competent state. Here we use the endogenous and closely spaced cysteine residues of the Pet toxin from enteroaggregative E. coli to investigate the effect of disulfide-bond-induced folding on translocation of an autotransporter passenger domain. We reveal that rigid structural elements within disulfide-bonded segments are resistant to autotransporter-mediated secretion. We define the size limit of disulfide-bonded segments tolerated by the autotransporter system demonstrating that, when present, cysteine pairs are intrinsically closely spaced to prevent congestion of the translocator pore by large disulfide-bonded regions. These latter data strongly support the hairpin mode of autotransporter biogenesis.

AB - Autotransporters are a super-family of virulence factors typified by a channel-forming C-terminus that facilitates translocation of the functional N-terminal passenger domain across the outer membrane of Gram-negative bacteria. This final step in the secretion of autotransporters requires a translocation-competent conformation for the passenger domain that differs markedly from the structure of the fully folded secreted protein. The nature of the translocation-competent conformation remains controversial, in particular whether the passenger domain can adopt secondary structural motifs, such as disulfide-bonded segments, while maintaining a secretion competent state. Here we use the endogenous and closely spaced cysteine residues of the Pet toxin from enteroaggregative E. coli to investigate the effect of disulfide-bond-induced folding on translocation of an autotransporter passenger domain. We reveal that rigid structural elements within disulfide-bonded segments are resistant to autotransporter-mediated secretion. We define the size limit of disulfide-bonded segments tolerated by the autotransporter system demonstrating that, when present, cysteine pairs are intrinsically closely spaced to prevent congestion of the translocator pore by large disulfide-bonded regions. These latter data strongly support the hairpin mode of autotransporter biogenesis.

U2 - 10.1074/jbc.M111.306118

DO - 10.1074/jbc.M111.306118

M3 - Article

C2 - 22006918

VL - 286

SP - 42283

EP - 42291

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 49

ER -