REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and virus strains
BL21(DE3) Competent Cells	Thermo Fisher Scientific	Cat# EC0114
Chemicals, peptides, and recombinant proteins
N-Dodecyl-b-D-Maltopyranoside (DDM), anagrade	Anatrace or Glycon	Cat# D310 or D97002
S-(2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl)methyl methanesulfonothioate (MTSSL)	Santa Cruz or Toronto Research Chemicals	Cat# 81213-52-7 or O875000
Glycerol	Fisher Scientific	G0650
Imidazole BioUltra	Sigma-Aldrich	56749
Sodium Chloride (NaCl)	Sigma-Aldrich	71383
LB Agar, Miller	Sigma-Aldrich	L3027
LB media, Miller	Sigma-Aldrich	L3152
TCEP-HCl	Thermo Scientific	20491
Phosphate buffered saline (PBS)	Sigma-Aldrich	P4417
Sodium Hydroxide (NaOH)	Fisher Scientific	BP359
Sodium phosphate dibasic dodecahydrate	Honeywell	04273
Sodium phosphate, monobasic dihydrate	Fisher Scientific	12645107
IPTG Dioxane Free	Formedium	367-93-1
Ampicillin	Formedium	69-52-3
D2O	Cambridge Isotope Laboratories	DLM-4-100
H2O (LC-MS Grade)	Supelco	1.15333
Acetonitrile	Supelco	1.00029
Formic acid	Supelco	5.33002
Critical commercial assays
Ni-NTA Agarose Resin	Invitrogen	Cat# R901-15
Superdex 200 Increase 10/300 GL column Superdex 200 16/600 GL column	Cytiva	Cat# 28-9909-44 Ca# 28-9893-35
Deposited data
MscL and MscL L89W HDX mass spectrometry data	Wang et al., 2022	ProteomeXchange PXD021983https://doi.org/10.1016/j.str.2021.12.004
MscL (various) mutants ESEEM data	Wang et al., 2022	http://archive.researchdata.leeds.ac.uk/777/https://doi.org/10.1016/j.str.2021.12.004
Oligonucleotides
Primer: MscL N13C Forward CGCGGTTGTATTGTTGACTTGGCGGT	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL N13C Reverse CAACAATACAACCGCGAGCCAGGAATTC	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL L17C Forward TGACTGCGCGGTTGCGGTTGTCATTGG	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL L17C Reverse CCGCGCAGTCAACAATATTACCGCGAGC	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL V21C Forward TTGCGTGTGTCATTGGTACCGCGTTTACCG	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL V21C Reverse CAATGACACACGCAACCGCCAAGTCAACAAT	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL V71C Forward GATTTGAATTGCCTGCTGAGCGCCGCTATTAAC	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL V71C Reverse GCAGGCAATTCAAATCGATGGTCTGACCACC	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL S74C Forward CTGCTGTGCGCCGCTATTAACTTC	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primer: MscL S74C Reverse GGCGCACAGCAGGACATTCAAATC	Wang et al., 2022	https://doi.org/10.1016/j.str.2021.12.004
Primers: remaining primers have been reported	Kapsalis et al. (2019)	https://doi.org/10.1038/s41467-019-12591-x
Recombinant DNA
Plasmid: pJ411:140126-TbMscL	Kapsalis et al. (2019)	https://doi.org/10.1038/s41467-019-12591-x
Software and algorithms
PLGS (v3.0.2)	Waters	https://www.waters.com/waters/en_GB/ProteinLynx-Global-SERVER-%28PLGS%29/nav.htm?locale=en_GB&cid=513821
DynamX (v3.0.0)	Waters	https://www.waters.com/waters/library.htm?locale=en_US&lid=134832928
PAVED	Cornwell et al. (2018)	https://doi.org/10.1007/s13361-018-2067-y
Deuteros	Lau et al. (2019)	https://github.com/andymlau/Deuteros_2.0
MATLAB Curve Fitting Toolbox	N/A	https://uk.mathworks.com/products/curvefitting.ht
Xepr Software	Bruker	https://www.bruker.com/en/products-and-solutions/mr/epr-instruments/epr-software.html
Other
Vivaspin-2 (100 kDa MWCO) Concentrator	Sartorius	Cat# VS0241