Browsing by Author "Selkrig, J"

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    Reconstitution of a nanomachine driving the assembly of proteins into bacterial outer membranes
    (Macmillan Publishers Limited., 2014-10-24) Shen, HH; Leyton, DL; Shiota, T; Belousoff, MJ; Noinaj, N; Lu, J; Holt, SA; Tan, K; Selkrig, J; Webb, CT; Buchanan, SK; Martin, LL; Lithgow, T
    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 Å, 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. © Macmillan Publishers Limited