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The organic anion transporter SLCO 2A1 constitutes the core component of the Maxi‐Cl channel
Author(s) -
Sabirov Ravshan Z,
Merzlyak Petr G,
Okada Toshiaki,
Islam Md Rafiqul,
Uramoto Hiromi,
Mori Tomoko,
Makino Yumiko,
Matsuura Hiroshi,
Xie Yu,
Okada Yasunobu
Publication year - 2017
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.15252/embj.201796685
Subject(s) - biology , transporter , component (thermodynamics) , core (optical fiber) , organic anion transporter 1 , channel (broadcasting) , microbiology and biotechnology , computational biology , biophysics , biochemistry , gene , computer science , computer network , telecommunications , physics , thermodynamics
The maxi‐anion channels ( MAC s) are expressed in cells from mammals to amphibians with ~60% exhibiting a phenotype called Maxi‐Cl. Maxi‐Cl serves as the most efficient pathway for regulated fluxes of inorganic and organic anions including ATP . However, its molecular entity has long been elusive. By subjecting proteins isolated from bleb membranes rich in Maxi‐Cl activity to LC ‐ MS / MS combined with targeted si RNA screening, CRISPR /Cas9‐mediated knockout, and heterologous overexpression, we identified the organic anion transporter SLCO 2A1, known as a prostaglandin transporter ( PGT ), as a key component of Maxi‐Cl. Recombinant SLCO 2A1 exhibited Maxi‐Cl activity in reconstituted proteoliposomes. When SLCO 2A1, but not its two disease‐causing mutants, was heterologously expressed in cells which lack endogenous SLCO 2A1 expression and Maxi‐Cl activity, Maxi‐Cl currents became activated. The charge‐neutralized mutant became weakly cation‐selective with exhibiting a smaller single‐channel conductance. Slco2a1 silencing in vitro and in vivo, respectively, suppressed the release of ATP from swollen C127 cells and from Langendorff‐perfused mouse hearts subjected to ischemia–reperfusion. These findings indicate that SLCO 2A1 is an essential core component of the ATP ‐conductive Maxi‐Cl channel.