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

36 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

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