1.A.114. The Proton-activated Chloride Channel (PACC) Family Severe local acidosis causes tissue damage and pain, and is one of the hallmarks of many diseases including ischemia, cancer, and inflammation. Yang et al. 2019 performed an unbiased RNA interference screen and identified PAC (TMEM206) as being essential for the widely observed proton-activated Cl^- (PAC) currents (I _Cl,H). Overexpression of human PAC in PAC knockout cells generated I _Cl,H with the same characteristics as the endogenous ones. Zebrafish PAC encodes a PAC channel with distinct properties. Knockout of mouse Pac abolished I _Cl,H in neurons and attenuated brain damage after ischemic stroke. The wide expression of PAC suggests a broad role for this conserved Cl^- channel family in physiological and pathological processes associated with acidic pH (Yang et al. 2019). It is involved in lysosome function, hypoxia adaption, stroke, and carcinogenesis (Cai et al. 2021). Ion permeation is controlled by hydrophobic residues and proton binding (Cai et al. 2021). Downregulation of TMEM206 inhibits malignant properties of human osteosarcoma cells (Peng et al. 2021). The plasma membrane inner leaflet PI(4,5)P₂ is essential for the activation of proton-activated chloride channels (Ko et al. 2024). Proton-activated chloride channels govern phagosome-mediated antibacterial immunity in peritoneal macrophages (Cheng et al. 2025). Ullrich et al. 2019 used a genome-wide siRNA screen to molecularly identify the widely expressed acid-sensitive outwardly-rectifying anion channel PAORAC/ASOR. ASOR is formed by TMEM206 proteins which display two TMSs and are expressed at the plasma membrane. Ion permeation-changing mutations along the length of TMS2 and at the end of TMS1 suggest that these segments line ASOR's pore. TMEM206 has orthologs in probably all vertebrates, but possibly not in other orgamism. Currents from evolutionarily distant orthologs share activation by protons, a feature essential for ASOR's role in acid-induced cell death (Ullrich et al. 2019). Molecular determinants of pH sensing have been determined, revealing that distinct pH-sensing and gating mechanisms are operative (Osei-Owusu et al. 2022). Phosphatidylinositol (4,5)-bisphosphate (PIP(2)) strongly inhibits the PAC channel by stabilizing the channel in a desensitized-like conformation (Mihaljević et al. 2023). PAC is active across a wide range of mammalian cells and is involved in acid-induced cell death and tissue injury. Ruan et al. 2020 presented two cryo-EM structures of human PAC in a high-pH resting closed state and a low-pH proton-bound non-conducting state. PAC is a trimer in which each subunit consists of a transmembrane domain (TMD), which is formed of two helices (TMS1 and TMS2), and an extracellular domain (ECD). Upon a decrease of pH from 8 to 4, a conformational change in the ECD-TMD interface and the TMD is observed. The rearrangement of the ECD-TMD interface is characterized by the movement of the histidine 98 residue, which is, after acidification, decoupled from the resting position and inserted into an acidic pocket that is about 5 Å away. Within the TMD, TMS1 undergoes a rotational movement, switching its interaction partner from its cognate TMS2 to the adjacent TMS2. The anion selectivity of PAC is determined by the positively charged lysine 319 residue in TMS2, and replacing lysine 319 with a glutamate residue converts PAC to a cation-selective channel (Ruan et al. 2020). ASOR (TMEM206/PAC) permeates anions across cellular membranes in response to acidification, thereby enhancing acid-induced cell death and regulating endocytosis. Wang et al. 2022 reconstituted function from purified protein and presented a 3.1-Å-resolution cryoEM structure of human ASOR at acidic pH in an activated conformation. The work contextualizes a previous acidic pH structure as a desensitized conformation. Combined with electrophysiological studies and high-resolution structures of resting and desensitized states, the work reveals mechanisms of proton sensing and ion pore gating. Clusters of extracellular acidic residues function as pH sensors and coalesce when protonated. Ensuing conformational changes induce metamorphosis of transmembrane helices to fashion an ion conduction pathway unique to the activated conformation. The studies identify a new paradigm of channel gating in this ubiquitous ion channel (Wang et al. 2022). Proton-activated chloride (PAC) channels play roles in regulating the pH and size of organelles in the endocytic pathway, and is also involved in acid-induced cell death. PAC (also known as TMEM206/ASOR) undergoes pH-dependent desensitization upon prolonged acid exposure (Osei-Owusu et al. 2022). Residues involved in desensitization have been identified.
References:
Cai, R., J. Tang, and X.Z. Chen. (2021). Ion permeation controlled by hydrophobic residues and proton binding in the proton-activated chloride channel. iScience 24: 103395.
Cheng, H.Y., J. Chu, N. Limjunyawong, J. Chen, Y. Ye, K.H. Chen, N. Koylass, S. Sun, X. Dong, and Z. Qiu. (2025). Proton-activated chloride channel governs phagosome-mediated antibacterial immunity in peritoneal macrophages. J Exp Med 222:.
Deng, Z., Y. Zhao, J. Feng, J. Zhang, H. Zhao, M.J. Rau, J.A.J. Fitzpatrick, H. Hu, and P. Yuan. (2021). Cryo-EM structure of a proton-activated chloride channel TMEM206. Sci Adv 7:.
Ko, W., E. Lee, J.E. Kim, H.H. Lim, and B.C. Suh. (2024). The plasma membrane inner leaflet PI(4,5)P is essential for the activation of proton-activated chloride channels. Nat Commun 15: 7008.
Maeda, S.I., R. Aoba, Y. Nishino, and T. Omata. (2019). A novel bacterial nitrate transporter composed of small transmembrane proteins. Plant Cell Physiol. [Epub: Ahead of Print]
Mihaljević, L., Z. Ruan, J. Osei-Owusu, W. Lü, and Z. Qiu. (2023). Inhibition of the proton-activated chloride channel PAC by PIP. Elife 12:.
Osei-Owusu, J., E. Kots, Z. Ruan, L. Mihaljević, K.H. Chen, A. Tamhaney, X. Ye, W. Lü, H. Weinstein, and Z. Qiu. (2022). Molecular determinants of pH sensing in the proton-activated chloride channel. Proc. Natl. Acad. Sci. USA 119: e2200727119.
Osei-Owusu, J., Z. Ruan, L. Mihaljević, D.S. Matasic, K.H. Chen, W. Lü, and Z. Qiu. (2022). Molecular mechanism underlying desensitization of the proton-activated chloride channel PAC. Elife 11:.
Peng, F., H. Li, J. Li, and Z. Wang. (2021). Downregulation of the Proton-Activated Cl^- Channel TMEM206 Inhibits Malignant Properties of Human Osteosarcoma Cells. Oxid Med Cell Longev 2021: 3672112.
Pissas, K.P., S. Gründer, and Y. Tian. (2024). Functional expression of the proton sensors ASIC1a, TMEM206, and OGR1 together with BK channels is associated with cell volume changes and cell death under strongly acidic conditions in DAOY medulloblastoma cells. Pflugers Arch 476: 923-937.
Ruan, Z., J. Osei-Owusu, J. Du, Z. Qiu, and W. Lü. (2020). Structures and pH-sensing mechanism of the proton-activated chloride channel. Nature. [Epub: Ahead of Print]
Ullrich, F., S. Blin, K. Lazarow, T. Daubitz, J.P. von Kries, and T.J. Jentsch. (2019). Identification of TMEM206 proteins as pore of PAORAC/ASOR acid-sensitive chloride channels. Elife 8:.
Wang, C., M.M. Polovitskaya, B.D. Delgado, T.J. Jentsch, and S.B. Long. (2022). Gating choreography and mechanism of the human proton-activated chloride channel ASOR. Sci Adv 8: eabm3942.
Xu, X.Y., F.F. Zhang, J. Gan, Z.S. Shen, Q. Guo, Y. Teng, J.W. Ji, J.L. Cao, Q.Y. Tang, and Z. Zhang. (2022). Identification of the Acid-Sensitive Site Critical for Chloral Hydrate (CH) Activation of the Proton-Activated Chloride Channel. J. Neurosci. [Epub: Ahead of Print]
Xue, P., W. Zhang, M. Shen, J. Yang, J. Chu, S. Wang, M. Wan, J. Zheng, Z. Qiu, and X. Cao. (2024). Proton-activated chloride channel increases endplate porosity and pain in a mouse spine degeneration model. J Clin Invest 134:.
Yang, J., J. Chen, M. Del Carmen Vitery, J. Osei-Owusu, J. Chu, H. Yu, S. Sun, and Z. Qiu. (2019). PAC, an evolutionarily conserved membrane protein, is a proton-activated chloride channel. Science 364: 395-399.
Examples:
TC#	Name	Organismal Type	Example
1.A.114.1.1	Plasma membrane proton-activated chloride channel, PACC1, or acid-sensitive outwardly-rectifying anion channel PAORAC/ASOR or TMEM206, of 350 aas and 2 TMSs. Ion permeation-changing mutations along the length of TMS2 and at the end of TMS1 suggest that these segments line the pore. TMEM206 probably has orthologs in all vertebrates (Ullrich et al. 2019). Knockout of mouse Pac abolished I _Cl,H in neurons and attenuated brain damage after ischemic stroke (Yang et al. 2019). The cryoEM structure (3.1 Å resolution) in active and desensitized states has been determined (Wang et al. 2022). The acid-sensitive site critical for chloral hydrate activation of the proton-activated chloride channel has been identified (Xu et al. 2022). The molecular mechanism underlying desensitization of the proton-activated chloride channel, PAC, has been examined (Osei-Owusu et al. 2022). PACC1 increases endplate porosity and pain in a mouse spine degeneration model (Xue et al. 2024).		PACC1 of Homo sapiens

1.A.114.1.2	Proton-activated chloride channel of 298 aas and 2 TMSs. It mediates import of chloride ion in response to extracellular acidic pH (Yang et al. 2019) and displays channel activity with kinetic properties distinct from that of the human ortholog.		TMEM206 of Danio rerio (Zebrafish) (Brachydanio rerio)

1.A.114.1.3	TMEM206 of 469 aas and 2 TMSs		TMEM206 of Amphimedon queenslandica

1.A.114.1.4	TMEM206-like protein of 382 aas and 2 TMSs.		TMEM206 of Saccoglossus kowalevskii

1.A.114.1.5	TMEM206 of 254 aas and 2 TMSs.		TMEM206 of Callorhinchus milii

1.A.114.1.6	Proton-activated chloride channel of 351 aas and 2 TMSs near the N- and C-termini. TMEM206 is an evolutionarily conserved chloride channel that underlies ubiquitously expressed, proton-activated, outwardly rectifying, anion currents. Deng et al. 2021 reported the cryo-EM structure of pufferfish TMEM206, which forms a trimeric channel, with 6 TMSs, 2 per subunit, each with a large extracellular domain. An ample vestibule in the extracellular region is accessible laterally from the three side portals. The central pore contains multiple constrictions; a conserved lysine residue near the cytoplasmic end of the inner helix forms the presumed chloride ion selectivity filter. The core structure and assembly closely resemble those of the epithelial sodium channel/degenerin family of sodium channels that seem to be unrelated in amino acid sequence and conduct cations instead of anions. Together with electrophysiology, this structure provides insights into ion conduction and gating for these chloride channels (Deng et al. 2021).		TMEM206 of Tetraodon nigroviridis (Spotted green pufferfish) (Chelonodon nigroviridis)