Darobactin B Stabilises a Lateral-Closed Conformation of the BAM Complex in E. coli Cells.
BAM
Cryoem
Darobactin
In-Cell EPR
PELDOR
Journal
Angewandte Chemie (International ed. in English)
ISSN: 1521-3773
Titre abrégé: Angew Chem Int Ed Engl
Pays: Germany
ID NLM: 0370543
Informations de publication
Date de publication:
21 08 2023
21 08 2023
Historique:
received:
19
12
2022
medline:
18
8
2023
pubmed:
10
5
2023
entrez:
10
5
2023
Statut:
ppublish
Résumé
The β-barrel assembly machinery (BAM complex) is essential for outer membrane protein (OMP) folding in Gram-negative bacteria, and represents a promising antimicrobial target. Several conformational states of BAM have been reported, but all have been obtained under conditions which lack the unique features and complexity of the outer membrane (OM). Here, we use Pulsed Electron-Electron Double Resonance (PELDOR, or DEER) spectroscopy distance measurements to interrogate the conformational ensemble of the BAM complex in E. coli cells. We show that BAM adopts a broad ensemble of conformations in the OM, while in the presence of the antibiotic darobactin B (DAR-B), BAM's conformational equilibrium shifts to a restricted ensemble consistent with the lateral closed state. Our in-cell PELDOR findings are supported by new cryoEM structures of BAM in the presence and absence of DAR-B. This work demonstrates the utility of PELDOR to map conformational changes in BAM within its native cellular environment.
Identifiants
pubmed: 37162386
doi: 10.1002/anie.202218783
doi:
Substances chimiques
Escherichia coli Proteins
0
darobactin
0
Bacterial Outer Membrane Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e202218783Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/S018069/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/W019795/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/M011151/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/P018491/1
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 108466/Z/15/Z
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 221524/Z/20/Z
Pays : United Kingdom
Organisme : Wellcome Trust
ID : 222373/Z/21/Z
Pays : United Kingdom
Informations de copyright
© 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.
Références
Angew Chem Int Ed Engl. 2009;48(16):2904-6
pubmed: 19294709
Molecules. 2019 Dec 06;24(24):
pubmed: 31817785
Proc Natl Acad Sci U S A. 2012 Oct 2;109(40):E2675-82
pubmed: 23012406
Biochemistry. 2018 Feb 13;57(6):1045-1053
pubmed: 29338257
Nature. 2020 Jul;583(7816):473-478
pubmed: 32528179
Angew Chem Int Ed Engl. 2016 Sep 12;55(38):11538-42
pubmed: 27511025
Elife. 2021 Jul 12;10:
pubmed: 34251336
J Magn Reson. 2015 Mar;252:187-98
pubmed: 25701439
Nat Struct Mol Biol. 2013 Feb;20(2):215-21
pubmed: 23334289
Nat Commun. 2021 Dec 8;12(1):7131
pubmed: 34880256
Curr Opin Struct Biol. 2022 Aug;75:102398
pubmed: 35667279
Nat Commun. 2019 Oct 10;10(1):4619
pubmed: 31601809
Angew Chem Int Ed Engl. 2023 Jan 9;62(2):e202214094
pubmed: 36308277
J Gen Physiol. 2021 Nov 1;153(11):
pubmed: 34529007
mBio. 2022 Oct 26;13(5):e0228622
pubmed: 36165532
Nature. 2019 Dec;576(7787):452-458
pubmed: 31645764
J Am Chem Soc. 2016 Feb 17;138(6):1844-7
pubmed: 26795032
Nat Commun. 2016 Sep 30;7:12865
pubmed: 27686148
Angew Chem Int Ed Engl. 2021 Aug 23;60(35):19155-19161
pubmed: 33844392
Annu Rev Phys Chem. 2012;63:419-46
pubmed: 22404592
Sci Adv. 2022 Oct 14;8(41):eabn6845
pubmed: 36223470
Proc Natl Acad Sci U S A. 2018 Apr 3;115(14):3692-3697
pubmed: 29555747
Angew Chem Int Ed Engl. 2019 Aug 19;58(34):11681-11685
pubmed: 31218813
Nat Struct Mol Biol. 2020 Sep;27(9):829-835
pubmed: 32719456
Nature. 2021 May;593(7857):125-129
pubmed: 33854236
Proc Natl Acad Sci U S A. 2022 Aug 23;119(34):e2206129119
pubmed: 35969794
Nat Commun. 2021 Sep 10;12(1):5387
pubmed: 34508071
Methods Enzymol. 2021;651:235-290
pubmed: 33888206
Angew Chem Int Ed Engl. 2016 Dec 19;55(51):15905-15909
pubmed: 27860003
Angew Chem Int Ed Engl. 2023 Aug 21;62(34):e202218783
pubmed: 37162386
Structure. 2014 Jul 8;22(7):1055-62
pubmed: 24980798
J Am Chem Soc. 2021 Nov 3;143(43):17875-17890
pubmed: 34664948
Annu Rev Microbiol. 2022 Sep 8;76:259-279
pubmed: 35650668
Proc Natl Acad Sci U S A. 2019 Oct 22;116(43):21748-21757
pubmed: 31591200
Annu Rev Microbiol. 2017 Sep 8;71:539-556
pubmed: 28886680
EMBO J. 2017 Oct 16;36(20):3062-3079
pubmed: 28864543
Nat Protoc. 2019 Aug;14(8):2344-2369
pubmed: 31278399
Proc Natl Acad Sci U S A. 2020 Jan 7;117(1):395-404
pubmed: 31862713
Nat Microbiol. 2022 Oct;7(10):1661-1672
pubmed: 36163500
Q Rev Biophys. 2007 Feb;40(1):1-53
pubmed: 17565764
Methods Enzymol. 2017;594:203-242
pubmed: 28779841
Angew Chem Int Ed Engl. 2022 Jan 10;61(2):e202113448
pubmed: 34761852
Chempluschem. 2020 Feb;85(2):353-372
pubmed: 31950648
Nature. 2016 Mar 3;531(7592):64-9
pubmed: 26901871
Nat Struct Mol Biol. 2016 Aug;23(8):744-51
pubmed: 27399258
Sci Adv. 2022 Nov 04;8(44):eadc9566
pubmed: 36322653
Microbiol Spectr. 2021 Dec 22;9(3):e0153521
pubmed: 34937193
Nature. 2019 Dec;576(7787):459-464
pubmed: 31747680
Nat Commun. 2021 Jul 7;12(1):4174
pubmed: 34234105
Biophys J. 2020 Jul 21;119(2):448-459
pubmed: 32621864
J Phys Chem B. 2022 Oct 6;126(39):7486-7494
pubmed: 36137278
Methods Enzymol. 2015;563:483-502
pubmed: 26478496
Angew Chem Int Ed Engl. 2015 May 18;54(21):6196-9
pubmed: 25826642