Table 3.

Available structures of LRRC8 in 2021

Structure	Functional state	PDB code	Species	Oligomeric state	Solubilization conditions	Freezing buffer conditions	Reference
LRRC8A 4.4 Å	Not commented on	6DJB	Homo sapiens	hexamer	20 mM Tris pH 8, 150 mM NaCl, 1% DMNG, 2 mg/mL iodoacetamide, and EDTA-free protease inhibitor cocktail	20 mM Tris pH 8, 150 mM NaCl, 0.05% digitonin, and EDTA-free protease inhibitor cocktail	Kefauver et al., 2018
LRRC8A 4.25 Å	Suggestion that the channel may be open but authors state they can not precisely define open/cosed states on the basis of this structure	5ZSU	Homo sapiens	hexamer	50 mM Tris, pH 8.0, 150 mM NaCl, 5 mM DTT, 1% digitonin	50 mM Tris, pH 8.0, 150 mM NaCl, 5 mM DTT, 0.1% digitonin	Kasuya et al., 2018
LRRC8A 4.18 Å	Apo-LRRC8A in a constricted state	6O00	Mus musculus	hexamer	A 10%/2% solution of DDM/CHS was dissolved and clarified by bath sonication in 200 mM HEPES pH 8 prior to addition to the solubilisation buffer. 50 mM HEPES, 150 mM KCl, 1 mM EDTA, 1% DDM, 0.2% CHS, final pH 7.4.	20 mM HEPES, 150 mM KCl, 1 mM EDTA pH 7.4, POPC, MSP2N2	Kern et al., 2019
LRRC8A-refined against 5.01 Å map	Authors suggest that the high salt conditions in the freezing buffer favor a closed conformation	6G9L	Mus musculus	hexamer	25 mM Tris-HCl, pH 8.5, 250 mM NaCl, 3% digitonin,50 μg ml−1 DNase, 50 μg ml−1 RNase A and protease inhibitors	25 mM Tris-HCl pH 8.5, 250 mM NaCl, 0.12% digitonin	Deneka and Sawicka et al., 2018
LRRC8A-refined against 4.25 Å map	Authors suggest that the high salt conditions in the freezing buffer favor a closed conformation	6G9O	Mus musculus	hexamer	25 mM Tris-HCl, pH 8.5, 250 mM NaCl, 3% digitonin,50 μg ml−1 DNase, 50 μg ml−1 RNase A and protease inhibitors	25 mM Tris-HCl pH 8.5, 250 mM NaCl, 0.12% digitonin	Deneka and Sawicka et al., 2018
LRRC8A-pore domain 3.66 Å	Authors suggest that the high salt conditions in the freezing buffer favor a closed conformation	6G8Z	Mus musculus	hexamer	25 mM Tris-HCl, pH 8.5, 250 mM NaCl, 3% digitonin,50 μg ml−1 DNase, 50 μg ml−1 RNase A and protease inhibitors	25 mM Tris-HCl pH 8.5, 250 mM NaCl, 0.12% digitonin	Deneka and Sawicka et al., 2018
LRRC8A-LRR domain 1.80 Å (X-ray structure)	n/a	6FNW	Mus musculus	protomer	10 mM Tris pH 9.4, 200 mM NaCl, 2% DDM, 50 μg ml−1 DNase, protease inhibitors	The protein was polished on SEC in 10 mM Tris pH 9.4, 200 mM NaCl, 0.1% CHAPS and then supplemented with 0.5% CHAPS and 1 mM tris(2-carboxyethyl)phosphine prior to crystallization in 0.1 M Bis-Tris propane pH 8.5, 0.2 M sodium malonate and 20% PEG3350	Deneka and Sawicka et al., 2018
LRRC8A-DCPIB 3.21 Å	Constricted state	6NZW	Mus musculus	hexamer	A 10%/2% solution of DDM/CHS was dissolved and clarified by bath sonication in 200 mM HEPES pH 8 prior to addition to the solubilisation buffer. 50 mM HEPES, 150 mM KCl, 1 mM EDTA, 1% DDM, 0.2% CHS, final pH 7.4.	20 mM HEPES, 150 mM KCl, 1 mM EDTA pH 7.4, POPC, MSP1E3D1. 100 μM of DCPIB was added prior to freezing.	Kern et al., 2019
LRRC8A-DCPIB 3.32 Å	Expanded state	6NZZ	Mus musculus	hexamer	A 10%/2% solution of DDM/CHS was dissolved and clarified by bath sonication in 200 mM HEPES pH 8 prior to addition to the solubilisation buffer. 50 mM HEPES, 150 mM KCl, 1 mM EDTA, 1% DDM, 0.2% CHS, final pH 7.4.	20 mM HEPES, 150 mM KCl, 1 mM EDTA pH 7.4, POPC, MSP1E3D1. 100 μM of DCPIB was added prior to freezing.	Kern et al., 2019
LRRC8D 4.36 Å	Undetermined. Some suggestion that the channel may be in an open conformation	6M04	Homo sapiens	hexamer	50 mM Tris, pH 8.0, 150 mM NaCl, 5 mM DTT, 1% digitonin	50 mM Tris, pH 8.0, 150 mM NaCl, 5 mM DTT, and 0.1% digitonin	Nakamura et al., 2020
LRRC8A/C heteromer	n/a	n/a (7.94 Å map)	Mus musculus	hexamer	25 mM Tris-HCl, pH 8.5, 250 mM NaCl, 3% digitonin,50 μg ml−1 DNase, 50 μg ml−1 RNase A and protease inhibitors	25 mM Tris-HCl pH 8.5, 250 mM NaCl, 0.12% digitonin	Deneka and Sawicka et al., 2018