Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes

Hsin-Hui Shen, Denisse L Leyton, Takuya Shiota, Matthew J Belousoff, Nicholas Noinaj, Jingxiong Lu, Stephen A Holt, Kher Shing Tan, Joel Pearson Selkrig, Chaille T Webb, Susan K Buchanan, Lisandra L Martin, Trevor J Lithgow

Research output: Contribution to journalArticleResearchpeer-review

35 Citations (Scopus)

Abstract

In biological membranes, various protein secretion devices function as nanomachines, and measuring the internal movements of their component parts is a major technological challenge. The translocation and assembly module (TAM) is a nanomachine required for virulence of bacterial pathogens. We have reconstituted a membrane containing the TAM onto a gold surface for characterization by quartz crystal microbalance with dissipation (QCM-D) and magnetic contrast neutron reflectrometry (MCNR). The MCNR studies provided structural resolution down to 1 A, enabling accurate measurement of protein domains projecting from the membrane layer. Here we show that dynamic movements within the TamA component of the TAM are initiated in the presence of a substrate protein, Ag43, and that these movements recapitulate an initial stage in membrane protein assembly. The reconstituted system provides a powerful new means to study molecular movements in biological membranes, and the technology is widely applicable to studying the dynamics of diverse cellular nanomachines.
Original languageEnglish
Pages (from-to)1 - 10
Number of pages10
JournalNature Communications
Volume5
Issue number5078
DOIs
Publication statusPublished - 2014

Cite this

Shen, Hsin-Hui ; Leyton, Denisse L ; Shiota, Takuya ; Belousoff, Matthew J ; Noinaj, Nicholas ; Lu, Jingxiong ; Holt, Stephen A ; Tan, Kher Shing ; Selkrig, Joel Pearson ; Webb, Chaille T ; Buchanan, Susan K ; Martin, Lisandra L ; Lithgow, Trevor J. / Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes. In: Nature Communications. 2014 ; Vol. 5, No. 5078. pp. 1 - 10.
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abstract = "In biological membranes, various protein secretion devices function as nanomachines, and measuring the internal movements of their component parts is a major technological challenge. The translocation and assembly module (TAM) is a nanomachine required for virulence of bacterial pathogens. We have reconstituted a membrane containing the TAM onto a gold surface for characterization by quartz crystal microbalance with dissipation (QCM-D) and magnetic contrast neutron reflectrometry (MCNR). The MCNR studies provided structural resolution down to 1 A, enabling accurate measurement of protein domains projecting from the membrane layer. Here we show that dynamic movements within the TamA component of the TAM are initiated in the presence of a substrate protein, Ag43, and that these movements recapitulate an initial stage in membrane protein assembly. The reconstituted system provides a powerful new means to study molecular movements in biological membranes, and the technology is widely applicable to studying the dynamics of diverse cellular nanomachines.",
author = "Hsin-Hui Shen and Leyton, {Denisse L} and Takuya Shiota and Belousoff, {Matthew J} and Nicholas Noinaj and Jingxiong Lu and Holt, {Stephen A} and Tan, {Kher Shing} and Selkrig, {Joel Pearson} and Webb, {Chaille T} and Buchanan, {Susan K} and Martin, {Lisandra L} and Lithgow, {Trevor J}",
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Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes. / Shen, Hsin-Hui; Leyton, Denisse L; Shiota, Takuya; Belousoff, Matthew J; Noinaj, Nicholas; Lu, Jingxiong; Holt, Stephen A; Tan, Kher Shing; Selkrig, Joel Pearson; Webb, Chaille T; Buchanan, Susan K; Martin, Lisandra L; Lithgow, Trevor J.

In: Nature Communications, Vol. 5, No. 5078, 2014, p. 1 - 10.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes

AU - Shen, Hsin-Hui

AU - Leyton, Denisse L

AU - Shiota, Takuya

AU - Belousoff, Matthew J

AU - Noinaj, Nicholas

AU - Lu, Jingxiong

AU - Holt, Stephen A

AU - Tan, Kher Shing

AU - Selkrig, Joel Pearson

AU - Webb, Chaille T

AU - Buchanan, Susan K

AU - Martin, Lisandra L

AU - Lithgow, Trevor J

PY - 2014

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