Journal article
Structure and mechanism of a tripartite ATP-independent periplasmic TRAP transporter
JS Davies, MJ Currie, RA North, M Scalise, JD Wright, JM Copping, DM Remus, A Gulati, DR Morado, SA Jamieson, MC Newton-Vesty, GS Abeysekera, S Ramaswamy, R Friemann, S Wakatsuki, JR Allison, C Indiveri, D Drew, PD Mace, RCJ Dobson
Nature Communications | Published : 2023
Open access
Abstract
In bacteria and archaea, tripartite ATP-independent periplasmic (TRAP) transporters uptake essential nutrients. TRAP transporters receive their substrates via a secreted soluble substrate-binding protein. How a sodium ion-driven secondary active transporter is strictly coupled to a substrate-binding protein is poorly understood. Here we report the cryo-EM structure of the sialic acid TRAP transporter SiaQM from Photobacterium profundum at 2.97 Å resolution. SiaM comprises a “transport” domain and a “scaffold” domain, with the transport domain consisting of helical hairpins as seen in the sodium ion-coupled elevator transporter VcINDY. The SiaQ protein forms intimate contacts with SiaM to ext..
View full abstractGrants
Awarded by University of California
Funding Acknowledgements
This research was undertaken in part using the MX2 beamline at the Australian Synchrotron, part of the Australian Nuclear Science and Technology Organisation (ANSTO), and made use of the Australian Cancer Research Foundation (ACRF) detector as well as the SAXS beamline at the Australian Synchrotron, part of ANSTO. R.C.J.D., R.A.N., J.R.A., S.W., J.S.D. and M.J.C. acknowledge funding support from the Marsden Fund, managed by Royal Society Te Aparangi (contract UOC1506) and the Biomolecular Interaction Centre (UC). R.C.J.D., J.S.D. and M.J.C. also acknowledge the following for funding support, in part: (1) the Ministry of Business, Innovation and Employment Smart Ideas grant (contract UOCX1706); (2) the Maurice Wilkins Centre flexible research grant; and (3) the Australian Institute of Nuclear Science and Engineering (AINSE Ltd) and ANSTO for a Postgraduate Research Award. R.A.N. acknowledges the Canterbury Medical Research Fund (contract CMRF 08). J.R.A. acknowledges the Rutherford Discovery Fellowship, managed by the Royal Society Te Aparangi (contract 15-MAU-001/15-UOA-008). J.C. is supported by a University of Auckland Doctoral Scholarship. The authors acknowledge the facilities, and scientific and technical assistance from flow cytometry staff at Otago Micro and Nanoscale Imaging (OMNI), at the University of Otago. We thank Prof. Borries Demeler (University of Lethbridge, Canada) for help with AUC experiments. R.F. acknowledges the Swedish Governmental Agency for Innovation Systems (2017-00180), and the Centre for Antibiotic Resistance Research (CARe) at the University of Gothenburg. C.I. acknowledges the MIUR (Ministry of Education, University and Research) Italy for the support through the "SI.F.I.PA.CRO.DE. - Sviluppo e industrializzazione farmaci innovativi per terapia molecolare personalizzata PA.CRO.DE." (PON ARS01_00568). D.D. acknowledges funding from the Knut and Alice Wallenberg Foundation. Cryo-EM data were collected at the Cryo-EM Swedish National Facility funded by the Knut and Alice Wallenberg Foundation, the Family Erling Persson and Kempe Foundations, SciLifeLab, Stockholm University and Umea University. We thank Aashish Manglik (University of California) and Andrew Kruse (Harvard University) for providing the original nanobody yeast-display library.