Autotransporter beta-domains have a specific function in protein secretion beyond outer-membrane targeting

A. Sauri, N. Oreshkova, Z. Soprova, W.S.P. Jong, M. Sani, P.J. Peters, J. Luirink, P. van Ulsen

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Autotransporters (ATs) of Gram-negative bacteria contain an N-proximal passenger domain that is transported to the extracellular milieu and a C-terminal β-domain that inserts into the outer membrane (OM) in a β-barrel conformation. This β-domain facilitates translocation of the passenger domain across the OM and has long been considered to be the translocation pore. However, available crystal structures of β-domains show that the β-barrel pore is too narrow for the observed transport of folded elements within the passenger domains. ATs have recently been shown to interact with the β-barrel assembly machinery. These findings questioned a direct involvement of the β-domain in passenger translocation and suggested that it may only target the passenger to the β-barrel assembly machinery pore. To address the function of the β-domain in more detail, we have replaced the β-domain of the Escherichia coli AT hemoglobin protease by β-domains originating from other OM proteins. Furthermore, we have modified the diameter of the β-domain pore. The mutant proteins were analyzed for their capacity to insert into the OM and for surface display of the passenger. Our results show that efficient passenger secretion requires a specific β-domain that not only functions as a targeting device but also is directly involved in the translocation of the passenger to the cell surface. © 2011 Elsevier Ltd. All rights reserved.
Original languageEnglish
Pages (from-to)553-67
JournalJournal of Molecular Biology
Volume412
DOIs
Publication statusPublished - 2011

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Membranes
Proteins
Mutant Proteins
Gram-Negative Bacteria
Membrane Proteins
Hemoglobins
Peptide Hydrolases
Escherichia coli
Equipment and Supplies
Type V Secretion Systems

Bibliographical note

1089-8638 Sauri, Ana Oreshkova, Nadia Soprova, Zora Jong, Wouter S P Sani, Musa Peters, Peter J Luirink, Joen van Ulsen, Peter Journal Article Research Support, Non-U.S. Gov't England J Mol Biol. 2011 Sep 30;412(4):553-67. doi: 10.1016/j.jmb.2011.07.035. Epub 2011 Jul 23.

Cite this

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title = "Autotransporter beta-domains have a specific function in protein secretion beyond outer-membrane targeting",
abstract = "Autotransporters (ATs) of Gram-negative bacteria contain an N-proximal passenger domain that is transported to the extracellular milieu and a C-terminal β-domain that inserts into the outer membrane (OM) in a β-barrel conformation. This β-domain facilitates translocation of the passenger domain across the OM and has long been considered to be the translocation pore. However, available crystal structures of β-domains show that the β-barrel pore is too narrow for the observed transport of folded elements within the passenger domains. ATs have recently been shown to interact with the β-barrel assembly machinery. These findings questioned a direct involvement of the β-domain in passenger translocation and suggested that it may only target the passenger to the β-barrel assembly machinery pore. To address the function of the β-domain in more detail, we have replaced the β-domain of the Escherichia coli AT hemoglobin protease by β-domains originating from other OM proteins. Furthermore, we have modified the diameter of the β-domain pore. The mutant proteins were analyzed for their capacity to insert into the OM and for surface display of the passenger. Our results show that efficient passenger secretion requires a specific β-domain that not only functions as a targeting device but also is directly involved in the translocation of the passenger to the cell surface. {\circledC} 2011 Elsevier Ltd. All rights reserved.",
author = "A. Sauri and N. Oreshkova and Z. Soprova and W.S.P. Jong and M. Sani and P.J. Peters and J. Luirink and {van Ulsen}, P.",
note = "1089-8638 Sauri, Ana Oreshkova, Nadia Soprova, Zora Jong, Wouter S P Sani, Musa Peters, Peter J Luirink, Joen van Ulsen, Peter Journal Article Research Support, Non-U.S. Gov't England J Mol Biol. 2011 Sep 30;412(4):553-67. doi: 10.1016/j.jmb.2011.07.035. Epub 2011 Jul 23.",
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Autotransporter beta-domains have a specific function in protein secretion beyond outer-membrane targeting. / Sauri, A.; Oreshkova, N.; Soprova, Z.; Jong, W.S.P.; Sani, M.; Peters, P.J.; Luirink, J.; van Ulsen, P.

In: Journal of Molecular Biology, Vol. 412, 2011, p. 553-67.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Autotransporter beta-domains have a specific function in protein secretion beyond outer-membrane targeting

AU - Sauri, A.

AU - Oreshkova, N.

AU - Soprova, Z.

AU - Jong, W.S.P.

AU - Sani, M.

AU - Peters, P.J.

AU - Luirink, J.

AU - van Ulsen, P.

N1 - 1089-8638 Sauri, Ana Oreshkova, Nadia Soprova, Zora Jong, Wouter S P Sani, Musa Peters, Peter J Luirink, Joen van Ulsen, Peter Journal Article Research Support, Non-U.S. Gov't England J Mol Biol. 2011 Sep 30;412(4):553-67. doi: 10.1016/j.jmb.2011.07.035. Epub 2011 Jul 23.

PY - 2011

Y1 - 2011

N2 - Autotransporters (ATs) of Gram-negative bacteria contain an N-proximal passenger domain that is transported to the extracellular milieu and a C-terminal β-domain that inserts into the outer membrane (OM) in a β-barrel conformation. This β-domain facilitates translocation of the passenger domain across the OM and has long been considered to be the translocation pore. However, available crystal structures of β-domains show that the β-barrel pore is too narrow for the observed transport of folded elements within the passenger domains. ATs have recently been shown to interact with the β-barrel assembly machinery. These findings questioned a direct involvement of the β-domain in passenger translocation and suggested that it may only target the passenger to the β-barrel assembly machinery pore. To address the function of the β-domain in more detail, we have replaced the β-domain of the Escherichia coli AT hemoglobin protease by β-domains originating from other OM proteins. Furthermore, we have modified the diameter of the β-domain pore. The mutant proteins were analyzed for their capacity to insert into the OM and for surface display of the passenger. Our results show that efficient passenger secretion requires a specific β-domain that not only functions as a targeting device but also is directly involved in the translocation of the passenger to the cell surface. © 2011 Elsevier Ltd. All rights reserved.

AB - Autotransporters (ATs) of Gram-negative bacteria contain an N-proximal passenger domain that is transported to the extracellular milieu and a C-terminal β-domain that inserts into the outer membrane (OM) in a β-barrel conformation. This β-domain facilitates translocation of the passenger domain across the OM and has long been considered to be the translocation pore. However, available crystal structures of β-domains show that the β-barrel pore is too narrow for the observed transport of folded elements within the passenger domains. ATs have recently been shown to interact with the β-barrel assembly machinery. These findings questioned a direct involvement of the β-domain in passenger translocation and suggested that it may only target the passenger to the β-barrel assembly machinery pore. To address the function of the β-domain in more detail, we have replaced the β-domain of the Escherichia coli AT hemoglobin protease by β-domains originating from other OM proteins. Furthermore, we have modified the diameter of the β-domain pore. The mutant proteins were analyzed for their capacity to insert into the OM and for surface display of the passenger. Our results show that efficient passenger secretion requires a specific β-domain that not only functions as a targeting device but also is directly involved in the translocation of the passenger to the cell surface. © 2011 Elsevier Ltd. All rights reserved.

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VL - 412

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JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

ER -