1. ABOUT THE DATASET -------------------- Title: Data for 'Outer membrane protein assembly mediated by BAM-SurA complexes'. [Dataset]. https://doi.org/10.5518/1460 Creator(s): Katherine L Fenn1*, Jim E Horne1*, Joel A Crossley1, Nils Böhringer2,3,4, Romany J Horne1, Till F. Schäberle2,3,4, Antonio N Calabrese1, Sheena E Radford1†, Neil A Ranson1† Organisation(s): [1] Astbury Centre for Structural Molecular Biology and School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, LS2 9JT, UK [2] Institute for Insect Biotechnology, Justus-Liebig-University Giessen, 35392 Giessen, Germany [3] German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, 35392 Giessen, Germany [4] Branch for Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), 35392 Giessen, Germany. Rights-holder(s):Unless otherwise stated, Copyright 2024 University of Leeds Publication Year: 2024 Description: The outer membrane is a formidable barrier that protects Gram-negative bacteria against environmental threats. Its integrity requires the correct folding and insertion of outer membrane proteins (OMPs) by the membrane-embedded β-barrel assembly machinery (BAM). Unfolded OMPs are delivered to BAM by the periplasmic chaperone SurA, but how SurA and BAM work together to ensure successful OMP delivery and folding remains unclear. Here, we present single-molecule Förster resonance energy transfer (smFRET) data of SurA and its interaction with the BAM alongside two alphafold predictions of the formed complex. Cite as: Katherine L Fenn*, Jim E Horne*, Joel A Crossley, Nils Böhringer, Romany J Horne, Till F. Schäberle, Antonio N Calabrese, Sheena E Radford†, Neil A Ranson†. (2024): Data for 'Outer membrane protein assembly mediated by BAM-SurA complexes'. [2024]. https://doi.org/10.5518/1460 Contact: j.a.crossley@leeds.ac.uk, k.l.fenn@leeds.ac.uk 2. TERMS OF USE --------------- Copyright 2024 University of Leeds. Unless otherwise stated, this dataset is licensed under a Creative Commons Attribution 4.0 International Licence: https://creativecommons.org/licenses/by/4.0/. 3. PROJECT AND FUNDING INFORMATION ---------------------------------- CryoEM data were collected at the Astbury Biostructure Laboratory, funded by the University of Leeds and Wellcome (108466/Z/15/Z; 221524/Z/20/Z) and we thank facility staff for their technical input. The Mass Spectrometry Facility instrumentation used in this work was funded by Wellcome (223810/Z/21/Z). KLF and JEH acknowledge funding from the MRC (MR/P018491/1). KLF is funded by the BBSRC (BB/X015653/1) and JAC by BBSRC (BB/T008059/1). SER holds a Royal Society Professorial Fellowship (RSRP/R1/211057). ANC acknowledges the support of a Sir Henry Dale Fellowship jointly funded by Wellcome and the Royal Society (220628/Z/20/Z) and a University Academic Fellowship from the University of Leeds. TFS and NB acknowledge funding from the German Federal Ministry of Education and Research (BMBF, via grant GBi2S and German Centre for Infection Research (DZIF) 09.918). 4. CONTENTS ----------- SurA_Core-PPIase1.hdf5 SurA_Core-PPIase1_BAM.hdf5 SurA_Core-PPIase1_WEYIPNV.hdf5 SurA_Core-PPIase1_WEYIPNV_BAM.hdf5 SurA_Core-PPIase2.hdf5 SurA_Core-PPIase2_BAM.hdf5 SurA_Core-PPIase2_WEYIPNV.hdf5 SurA_Core-PPIase2_WEYIPNV_BAM.hdf5 BAM_SurA_AlphaFold2Multimer.pdb SurA_POTRA1-5_ColabFoldPrediction.pdb 5. METHODS ---------- Single-molecule Förster resonance energy transfer (smFRET) experiments were performed on a custom-built confocal epi-illuminated microscope in a standard inverted-stage configuration with a pulsed interleaved excitation regime. For 'SurA_POTRA1-5_ColabFoldPrediction.pdb' the sequence of mature SurA (residues 21-427) and BamA POTRA domains (residues 21-424) from E. coli K-12 were submitted to the Google Colab servers for AlphaFold2 from Google DeepMind and ColabFold accessed on 29/07/2021 (now at github.com/sokrypton/ColabFold). For 'BAM_SurA_AlphaFold2Multimer.pdb' we used Alphafold-Multimer (v2.1.0) installed on a local workstation to predict the structure of the BAM–SurA complex using the reduced databases as described at https://github.com/deepmind/alphafold.