Mechanical Unfolding of an Autotransporter Passenger Protein Reveals the Secretion Starting Point and Processive Transport Intermediates

M. Baclayon, P. van Ulsen, H. Mouhib, M. Hashemi Shabestari, T. Verzijden, S. Abeln, W.H. Roos, G.J.L. Wuite

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The backbone of secreted autotransporter passenger proteins generally attains a stable β-helical structure. The secretion of passengers across the outer membrane was proposed to be driven by sequential folding of this structure at the cell surface. This mechanism would require a relatively stable intermediate as starting point. Here, we investigated the mechanics of secreted truncated versions of the autotransporter hemoglobin protease (Hbp) of Escherichia coli using atomic force microscopy. The data obtained reveal a β-helical structure at the C terminus that is very stable. In addition, several other distinct metastable intermediates are found which are connected during unfolding by multiroute pathways. Computational analysis indicates that these intermediates correlate to the β-helical rungs in the Hbp structure which are clamped by stacked aromatic residues. Our results suggest a secretion mechanism that is initiated by a stable C-terminal structure and driven forward by several folding intermediates that build up the β-helical backbone.
Original languageEnglish
Pages (from-to)5710-5719
JournalACS Nano
Volume10
Issue number6
DOIs
Publication statusPublished - 2016

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protease
secretions
passengers
Hemoglobin
hemoglobin
folding
Hemoglobins
Peptide Hydrolases
proteins
Proteins
Escherichia
Escherichia coli
Atomic force microscopy
Mechanics
atomic force microscopy
membranes
Membranes
cells
Type V Secretion Systems

Cite this

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title = "Mechanical Unfolding of an Autotransporter Passenger Protein Reveals the Secretion Starting Point and Processive Transport Intermediates",
abstract = "The backbone of secreted autotransporter passenger proteins generally attains a stable β-helical structure. The secretion of passengers across the outer membrane was proposed to be driven by sequential folding of this structure at the cell surface. This mechanism would require a relatively stable intermediate as starting point. Here, we investigated the mechanics of secreted truncated versions of the autotransporter hemoglobin protease (Hbp) of Escherichia coli using atomic force microscopy. The data obtained reveal a β-helical structure at the C terminus that is very stable. In addition, several other distinct metastable intermediates are found which are connected during unfolding by multiroute pathways. Computational analysis indicates that these intermediates correlate to the β-helical rungs in the Hbp structure which are clamped by stacked aromatic residues. Our results suggest a secretion mechanism that is initiated by a stable C-terminal structure and driven forward by several folding intermediates that build up the β-helical backbone.",
author = "M. Baclayon and {van Ulsen}, P. and H. Mouhib and {Hashemi Shabestari}, M. and T. Verzijden and S. Abeln and W.H. Roos and G.J.L. Wuite",
year = "2016",
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language = "English",
volume = "10",
pages = "5710--5719",
journal = "ACS Nano",
issn = "1936-0851",
publisher = "American Chemical Society",
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Mechanical Unfolding of an Autotransporter Passenger Protein Reveals the Secretion Starting Point and Processive Transport Intermediates. / Baclayon, M.; van Ulsen, P.; Mouhib, H.; Hashemi Shabestari, M.; Verzijden, T.; Abeln, S.; Roos, W.H.; Wuite, G.J.L.

In: ACS Nano, Vol. 10, No. 6, 2016, p. 5710-5719.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Mechanical Unfolding of an Autotransporter Passenger Protein Reveals the Secretion Starting Point and Processive Transport Intermediates

AU - Baclayon, M.

AU - van Ulsen, P.

AU - Mouhib, H.

AU - Hashemi Shabestari, M.

AU - Verzijden, T.

AU - Abeln, S.

AU - Roos, W.H.

AU - Wuite, G.J.L.

PY - 2016

Y1 - 2016

N2 - The backbone of secreted autotransporter passenger proteins generally attains a stable β-helical structure. The secretion of passengers across the outer membrane was proposed to be driven by sequential folding of this structure at the cell surface. This mechanism would require a relatively stable intermediate as starting point. Here, we investigated the mechanics of secreted truncated versions of the autotransporter hemoglobin protease (Hbp) of Escherichia coli using atomic force microscopy. The data obtained reveal a β-helical structure at the C terminus that is very stable. In addition, several other distinct metastable intermediates are found which are connected during unfolding by multiroute pathways. Computational analysis indicates that these intermediates correlate to the β-helical rungs in the Hbp structure which are clamped by stacked aromatic residues. Our results suggest a secretion mechanism that is initiated by a stable C-terminal structure and driven forward by several folding intermediates that build up the β-helical backbone.

AB - The backbone of secreted autotransporter passenger proteins generally attains a stable β-helical structure. The secretion of passengers across the outer membrane was proposed to be driven by sequential folding of this structure at the cell surface. This mechanism would require a relatively stable intermediate as starting point. Here, we investigated the mechanics of secreted truncated versions of the autotransporter hemoglobin protease (Hbp) of Escherichia coli using atomic force microscopy. The data obtained reveal a β-helical structure at the C terminus that is very stable. In addition, several other distinct metastable intermediates are found which are connected during unfolding by multiroute pathways. Computational analysis indicates that these intermediates correlate to the β-helical rungs in the Hbp structure which are clamped by stacked aromatic residues. Our results suggest a secretion mechanism that is initiated by a stable C-terminal structure and driven forward by several folding intermediates that build up the β-helical backbone.

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