National Institutes of Health/National Institute of Diabetes and Digestive and Kidney Disease (NIH/NIDDK)
DK51610
米国
引用
ジャーナル: Elife / 年: 2023 タイトル: Cryo-EM structures of an LRRC8 chimera with native functional properties reveal heptameric assembly. 著者: Hirohide Takahashi / Toshiki Yamada / Jerod S Denton / Kevin Strange / Erkan Karakas / 要旨: Volume-regulated anion channels (VRACs) mediate volume regulatory Cl and organic solute efflux from vertebrate cells. VRACs are heteromeric assemblies of LRRC8A-E proteins with unknown ...Volume-regulated anion channels (VRACs) mediate volume regulatory Cl and organic solute efflux from vertebrate cells. VRACs are heteromeric assemblies of LRRC8A-E proteins with unknown stoichiometries. Homomeric LRRC8A and LRRC8D channels have a small pore, hexameric structure. However, these channels are either non-functional or exhibit abnormal regulation and pharmacology, limiting their utility for structure-function analyses. We circumvented these limitations by developing novel homomeric LRRC8 chimeric channels with functional properties consistent with those of native VRAC/LRRC8 channels. We demonstrate here that the LRRC8C-LRRC8A(IL1) chimera comprising LRRC8C and 25 amino acids unique to the first intracellular loop (IL1) of LRRC8A has a heptameric structure like that of homologous pannexin channels. Unlike homomeric LRRC8A and LRRC8D channels, heptameric LRRC8C-LRRC8A(IL1) channels have a large-diameter pore similar to that estimated for native VRACs, exhibit normal DCPIB pharmacology, and have higher permeability to large organic anions. Lipid-like densities are located between LRRC8C-LRRC8A(IL1) subunits and occlude the channel pore. Our findings provide new insights into VRAC/LRRC8 channel structure and suggest that lipids may play important roles in channel gating and regulation.
名称: Volume-regulated anion channel subunit LRRC8C,Volume-regulated anion channel subunit LRRC8A タイプ: protein_or_peptide / ID: 1 詳細: LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag.,LRRC8C chimera where ...詳細: LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag.,LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag.,LRRC8C chimera where residues 184-204 are swapped with LRRC8A residues 182-206. There are also C-terminal TEV protease recognition site and FLAG purification tag. コピー数: 7 / 光学異性体: LEVO