{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T06:07:08Z","timestamp":1790662028322,"version":"4.1.0"},"reference-count":112,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2022,11,29]],"date-time":"2022-11-29T00:00:00Z","timestamp":1669680000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Pew Charitable Foundation","award":["1R01 DK128592"],"award-info":[{"award-number":["1R01 DK128592"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Membranes"],"abstract":"<jats:p>Loop and thiazide diuretics have been cornerstones of clinical management of hypertension and fluid overload conditions for more than five decades. The hunt for their molecular targets led to the discovery of cation-chloride cotransporters (CCCs) that catalyze electroneutral movement of Cl\u2212 together with Na+ and\/or K+. CCCs consist of two 1 Na+-1 K+-2 Cl\u2212 (NKCC1-2), one 1 Na+-1 Cl\u2212 (NCC), and four 1 K+-1 Cl\u2212 (KCC1-4) transporters in human. CCCs are fundamental in trans-epithelia ion secretion and absorption, homeostasis of intracellular Cl\u2212 concentration and cell volume, and regulation of neuronal excitability. Malfunction of NKCC2 and NCC leads to abnormal salt and water retention in the kidney and, consequently, imbalance in electrolytes and blood pressure. Mutations in KCC2 and KCC3 are associated with brain disorders due to impairments in regulation of excitability and possibly cell volume of neurons. A recent surge of structures of CCCs have defined their dimeric architecture, their ion binding sites, their conformational changes associated with ion translocation, and the mechanisms of action of loop diuretics and small molecule inhibitors. These breakthroughs now set the stage to expand CCC pharmacology beyond loop and thiazide diuretics, developing the next generation of diuretics with improved potency and specificity. Beyond drugging renal-specific CCCs, brain-penetrable therapeutics are sorely needed to target CCCs in the nervous system for the treatment of neurological disorders and psychiatric conditions.<\/jats:p>","DOI":"10.3390\/membranes12121206","type":"journal-article","created":{"date-parts":[[2022,11,30]],"date-time":"2022-11-30T01:52:20Z","timestamp":1669773140000},"page":"1206","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Structural Pharmacology of Cation-Chloride Cotransporters"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8209-6015","authenticated-orcid":false,"given":"Yongxiang","family":"Zhao","sequence":"first","affiliation":[{"name":"Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84102, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erhu","family":"Cao","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT 84102, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1258\/acb.2011.011281","article-title":"Diuretics: A review","volume":"49","author":"Wile","year":"2012","journal-title":"Ann. Clin. Biochem."},{"key":"ref_2","unstructured":"Seldin, D.W.G.G.H. (1997). Diuretic Agents: Clinical Physiology and Pharmacology, Academic Press."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1002\/j.1552-4604.1981.tb05661.x","article-title":"Bumetanide the Way to Its Chemical-Structure","volume":"21","author":"Feit","year":"1981","journal-title":"J. Clin. Pharmacol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"87","DOI":"10.3181\/00379727-49-13474","article-title":"Effect of some sulfonamides on renal secretion","volume":"49","author":"Hober","year":"1942","journal-title":"Proc. Soc. Exp. Biol. Med."},{"key":"ref_5","first-page":"199","article-title":"Carbonic anhydrase inhibition. I. The pharmacology of diamox 2-acetylamino-1,3,4-thiadiazole-5-sulfonamide","volume":"95","author":"Maren","year":"1954","journal-title":"Bull. Johns. Hopkins. Hosp."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1111\/j.1749-6632.1958.tb46763.x","article-title":"The mechanism of action of chlorothiazide","volume":"71","author":"Beyer","year":"1958","journal-title":"Ann. New York Acad. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1695","DOI":"10.1055\/s-0028-1112283","article-title":"Experimental Studies and Clinical Trials of a New Diuretic","volume":"88","author":"Kleinfelder","year":"1963","journal-title":"Dtsch. Med. Wochenschr."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1130","DOI":"10.1093\/ajh\/hpw030","article-title":"Diuretics for Hypertension: A Review and Update","volume":"29","author":"Roush","year":"2016","journal-title":"Am. J. Hypertens."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1172\/JCI111420","article-title":"Sodium-Chloride Absorption by the Urinary-Bladder of the Winter Flounder\u2014A Thiazide-Sensitive, Electrically Neutral Transport-System","volume":"74","author":"Stokes","year":"1984","journal-title":"J. Clin. Investig."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"C243","DOI":"10.1152\/ajpcell.1987.253.2.C243","article-title":"Photolabeling of a 150-Kda (Na+K+Cl) Cotransport Protein from Dog Kidney with a Bumetanide Analog","volume":"253","author":"Haas","year":"1987","journal-title":"Am. J. Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2201","DOI":"10.1073\/pnas.91.6.2201","article-title":"Molecular cloning and functional expression of the bumetanide-sensitive Na-K-Cl cotransporter","volume":"91","author":"Xu","year":"1994","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2749","DOI":"10.1073\/pnas.90.7.2749","article-title":"Primary structure and functional expression of a cDNA encoding the thiazide-sensitive, electroneutral sodium-chloride cotransporter","volume":"90","author":"Gamba","year":"1993","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"16237","DOI":"10.1074\/jbc.271.27.16237","article-title":"Molecular cloning and functional expression of the K-Cl cotransporter from rabbit, rat, and human. A new member of the cation-chloride cotransporter family","volume":"271","author":"Gillen","year":"1996","journal-title":"J. Biol. Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"16245","DOI":"10.1074\/jbc.271.27.16245","article-title":"Molecular characterization of a putative K-Cl cotransporter in rat brain. A neuronal-specific isoform","volume":"271","author":"Payne","year":"1996","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"C1210","DOI":"10.1152\/ajpcell.1999.277.6.C1210","article-title":"Molecular cloning and functional characterization of KCC3, a new K-Cl cotransporter","volume":"277","author":"Race","year":"1999","journal-title":"Am. J. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"16355","DOI":"10.1074\/jbc.274.23.16355","article-title":"Cloning and characterization of KCC3 and KCC4, new members of the cation-chloride cotransporter gene family","volume":"274","author":"Mount","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"4544","DOI":"10.1073\/pnas.91.10.4544","article-title":"Alternatively Spliced Isoforms of the Putative Renal Na-K-Cl Cotransporter Are Differentially Distributed within the Rabbit Kidney","volume":"91","author":"Payne","year":"1994","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1152\/physrev.2000.80.1.211","article-title":"Sodium-potassium-chloride cotransport","volume":"80","author":"Russell","year":"2000","journal-title":"Physiol. Rev."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1152\/physrev.00011.2004","article-title":"Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters","volume":"85","author":"Gamba","year":"2005","journal-title":"Physiol. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"C693","DOI":"10.1152\/ajpcell.00350.2012","article-title":"Physiology of SLC12 transporters: Lessons from inherited human genetic mutations and genetically engineered mouse knockouts","volume":"304","author":"Gagnon","year":"2013","journal-title":"Am. J. Physiol.-Cell Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1002\/jcp.21814","article-title":"Molecular characterization of a human cation-Cl- cotransporter (SLC12A8A, CCC9A) that promotes polyamine and amino acid transport","volume":"220","author":"Daigle","year":"2009","journal-title":"J. Cell. Physiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1038\/s42255-019-0088-x","article-title":"Slc12a8 is a nicotinamide mononucleotide transporter","volume":"1","author":"Grozio","year":"2019","journal-title":"Nat. Metab."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1038\/s42255-019-0086-z","article-title":"Reply to: Absence of evidence that Slc12a8 encodes a nicotinamide mononucleotide transporter","volume":"1","author":"Grozio","year":"2019","journal-title":"Nat. Metab."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"32027","DOI":"10.1074\/jbc.M000108200","article-title":"Cloning and functional characterization of a cation-Cl- cotransporter-interacting protein","volume":"275","author":"Caron","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1016\/j.bbrc.2009.02.057","article-title":"CIP1 is an activator of the K+-Cl- cotransporter KCC2","volume":"381","author":"Wenz","year":"2009","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1146\/annurev.physiol.64.081501.155847","article-title":"Human and murine phenotypes associated with defects in cation-chloride cotransport","volume":"64","author":"Delpire","year":"2002","journal-title":"Annu. Rev. Physiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1016\/j.apsb.2020.09.009","article-title":"Regulatory control of the Na-Cl co-transporter NCC and its therapeutic potential for hypertension","volume":"11","author":"Koeners","year":"2021","journal-title":"Acta. Pharm. Sin. B"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"F991","DOI":"10.1152\/ajprenal.00432.2014","article-title":"Physiology and pathophysiology of the renal Na-K-2Cl cotransporter (NKCC2)","volume":"307","author":"Castrop","year":"2014","journal-title":"Am. J. Physiol. Ren. Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1038\/ng0696-183","article-title":"Bartter\u2019s syndrome, hypokalaemic alkalosis with hypercalciuria, is caused by mutations in the Na-K-2Cl cotransporter NKCC2","volume":"13","author":"Simon","year":"1996","journal-title":"Nat. Genet."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/ng0196-24","article-title":"Gitelman\u2019s variant of Bartter's syndrome, inherited hypokalaemic alkalosis, is caused by mutations in the thiazide-sensitive Na-Cl cotransporter","volume":"12","author":"Simon","year":"1996","journal-title":"Nat. Genet."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1007\/s10157-011-0489-y","article-title":"The role of the kidney in salt-sensitive hypertension","volume":"16","author":"Trepiccione","year":"2012","journal-title":"Clin. Exp. Nephrol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"859","DOI":"10.1007\/s40620-021-01083-1","article-title":"Genotype-phenotype correlation in Gordon\u2019s syndrome: Report of two cases carrying novel heterozygous mutations","volume":"35","author":"Anglani","year":"2022","journal-title":"J. Nephrol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1146\/annurev.physiol.70.113006.100651","article-title":"Molecular physiology of the WNK kinases","volume":"70","author":"Kahle","year":"2008","journal-title":"Annu. Rev. Physiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1038\/nature10814","article-title":"Mutations in kelch-like 3 and cullin 3 cause hypertension and electrolyte abnormalities","volume":"482","author":"Boyden","year":"2012","journal-title":"Nature"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1111\/j.1751-7176.2011.00512.x","article-title":"Thiazide and Loop Diuretics","volume":"13","author":"Sica","year":"2011","journal-title":"J. Clin. Hypertens."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"820","DOI":"10.1016\/j.neuron.2009.03.003","article-title":"Cation-Chloride Cotransporters and Neuronal Function","volume":"61","author":"Blaesse","year":"2009","journal-title":"Neuron"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1038\/nrn3819","article-title":"Cation-chloride cotransporters in neuronal development, plasticity and disease","volume":"15","author":"Kaila","year":"2014","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2358","DOI":"10.1046\/j.1460-9568.2002.02419.x","article-title":"Patterns of cation-chloride cotransporter expression during embryonic rodent CNS development","volume":"16","author":"Li","year":"2002","journal-title":"Eur. J. Neurosci."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4134","DOI":"10.1523\/JNEUROSCI.23-10-04134.2003","article-title":"Expression and function of chloride transporters during development of inhibitory neurotransmission in the auditory brainstem","volume":"23","author":"Balakrishnan","year":"2003","journal-title":"J. Neurosci."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Watanabe, M., Zhang, J., Mansuri, M.S., Duan, J., Karimy, J.K., Delpire, E., Alper, S.L., Lifton, R.P., Fukuda, A., and Kahle, K.T. (2019). Developmentally regulated KCC2 phosphorylation is essential for dynamic GABA-mediated inhibition and survival. Sci. Signal., 12.","DOI":"10.1126\/scisignal.aaw9315"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4574","DOI":"10.1093\/cercor\/bhv218","article-title":"Developmental Expression Patterns of KCC2 and Functionally Associated Molecules in the Human Brain","volume":"26","author":"Sedmak","year":"2016","journal-title":"Cereb. Cortex."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1038\/16697","article-title":"The K+\/Cl- co-transporter KCC2 renders GABA hyperpolarizing during neuronal maturation","volume":"397","author":"Rivera","year":"1999","journal-title":"Nature"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"515","DOI":"10.3389\/fncel.2019.00515","article-title":"Identification of KCC2 Mutations in Human Epilepsy Suggests Strategies for Therapeutic Transporter Modulation","volume":"13","author":"Duy","year":"2019","journal-title":"Front. Cell Neurosci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"8038","DOI":"10.1038\/ncomms9038","article-title":"Mutations in SLC12A5 in epilepsy of infancy with migrating focal seizures","volume":"6","author":"Stodberg","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.1212\/01.wnl.0000204181.31175.8b","article-title":"Novel truncating and missense mutations of the KCC3 gene associated with Andermann syndrome","volume":"67","author":"Uyanik","year":"2006","journal-title":"Neurology"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"832","DOI":"10.1016\/j.trechm.2021.05.004","article-title":"Cation-coupled chloride cotransporters: Chemical insights and disease implications","volume":"3","author":"Portioli","year":"2021","journal-title":"Trends. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1016\/j.tips.2021.09.005","article-title":"Pharmacological tools to target NKCC1 in brain disorders","volume":"42","author":"Savardi","year":"2021","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"310","DOI":"10.3389\/fnins.2019.00310","article-title":"Off-Label Use of Bumetanide for Brain Disorders: An Overview","volume":"13","author":"Kharod","year":"2019","journal-title":"Front. Neurosci.-Switz."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"108910","DOI":"10.1016\/j.neuropharm.2021.108910","article-title":"CNS pharmacology of NKCC1 inhibitors*","volume":"205","author":"Loscher","year":"2022","journal-title":"Neuropharmacology"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"107616","DOI":"10.1016\/j.yebeh.2020.107616","article-title":"The search for brain-permeant NKCC1 inhibitors for the treatment of seizures: Pharmacokinetic-pharmacodynamic modelling of NKCC1 inhibition by azosemide, torasemide, and bumetanide in mouse brain","volume":"114","author":"Hampel","year":"2021","journal-title":"Epilepsy Behav."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1038\/s43587-021-00122-7","article-title":"Experimental and real-world evidence supporting the computational repurposing of bumetanide for APOE4-related Alzheimer\u2019s disease","volume":"1","author":"Taubes","year":"2021","journal-title":"Nat. Aging"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"806","DOI":"10.1111\/epi.12620","article-title":"Pharmacotherapeutic targeting of cation-chloride cotransporters in neonatal seizures","volume":"55","author":"Puskarjov","year":"2014","journal-title":"Epilepsia"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"C324","DOI":"10.1152\/ajpcell.00566.2020","article-title":"Advances in the development of novel compounds targeting cation-chloride cotransporter physiology","volume":"320","author":"Delpire","year":"2021","journal-title":"Am. J. Physiol.-Cell Physiol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"2073","DOI":"10.1016\/j.chempr.2020.06.017","article-title":"Discovery of a Small Molecule Drug Candidate for Selective NKCC1 Inhibition in Brain Disorders","volume":"6","author":"Savardi","year":"2020","journal-title":"Chem"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"5383","DOI":"10.1073\/pnas.0812756106","article-title":"Small-molecule screen identifies inhibitors of the neuronal K-Cl cotransporter KCC2","volume":"106","author":"Delpire","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1524","DOI":"10.1038\/nm.3356","article-title":"Chloride extrusion enhancers as novel therapeutics for neurological diseases","volume":"19","author":"Gagnon","year":"2013","journal-title":"Nat. Med."},{"key":"ref_57","first-page":"224","article-title":"Treatment of congestive heart failure and hypertension: Clinical experience with hydrochlorothiazide","volume":"11","author":"Weiner","year":"1960","journal-title":"Am. Pract. Dig. Treat."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1640","DOI":"10.1136\/bmj.2.5425.1640","article-title":"Today\u2019s Drugs. Frusemide","volume":"2","author":"Ingram","year":"1964","journal-title":"Br. Med. J."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1136\/bmj.1.5794.211","article-title":"Bumetanide: Potent new \u201cloop\u201d diuretic","volume":"1","author":"Asbury","year":"1972","journal-title":"Br. Med. J."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1002\/j.1875-9114.1984.tb03348.x","article-title":"Piretanide: A loop-active diuretic. Pharmacology, therapeutic efficacy and adverse effects","volume":"4","author":"Marsh","year":"1984","journal-title":"Pharmacotherapy"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1038\/clpt.1987.131","article-title":"Clinical pharmacology of torasemide, a new loop diuretic","volume":"42","author":"Brater","year":"1987","journal-title":"Clin. Pharmacol. Ther."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1080\/00325481.1976.11714327","article-title":"Diuretic agents. Mechanisms of action and clinical uses","volume":"59","author":"Krumlovsky","year":"1976","journal-title":"Postgrad. Med."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"5072","DOI":"10.1074\/jbc.M508050200","article-title":"Decreased renal organic anion secretion and plasma accumulation of endogenous organic anions in OAT1 knock-out mice","volume":"281","author":"Eraly","year":"2006","journal-title":"J Biol. Chem."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"F867","DOI":"10.1152\/ajprenal.00528.2007","article-title":"Overlapping in vitro and in vivo specificities of the organic anion transporters OAT1 and OAT3 for loop and thiazide diuretics","volume":"294","author":"Vallon","year":"2008","journal-title":"Am. J. Physiol.-Ren."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/S0030-6665(20)30770-2","article-title":"Ototoxicity of loop diuretics","volume":"26","author":"Rybak","year":"1993","journal-title":"Otolaryngol. Clin. N. Am."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"C225","DOI":"10.1152\/ajpcell.00465.2019","article-title":"Cryo-EM structures of DrNKCC1 and hKCC1: A new milestone in the physiology of cation-chloride cotransporters","volume":"318","author":"Delpire","year":"2020","journal-title":"Am. J. Physiol. Cell. Physiol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"167056","DOI":"10.1016\/j.jmb.2021.167056","article-title":"High-Resolution Views and Transport Mechanisms of the NKCC1 and KCC Transporters","volume":"433","author":"Chew","year":"2021","journal-title":"J. Mol. Biol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.gendis.2015.02.007","article-title":"NKCC1 and NKCC2: The pathogenetic role of cation-chloride cotransporters in hypertension","volume":"2","author":"Orlov","year":"2015","journal-title":"Genes Dis."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"C1516","DOI":"10.1152\/ajpcell.1997.273.5.C1516","article-title":"Functional characterization of the neuronal-specific K-Cl cotransporter: Implications for [K+]o regulation","volume":"273","author":"Payne","year":"1997","journal-title":"Am. J. Physiol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"30326","DOI":"10.1074\/jbc.M003112200","article-title":"Functional comparison of the K+-Cl- cotransporters KCC1 and KCC4","volume":"275","author":"Mercado","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1016\/j.str.2009.02.009","article-title":"X-ray Structure of the C-Terminal Domain of a Prokaryotic Cation-Chloride Cotransporter","volume":"17","author":"Warmuth","year":"2009","journal-title":"Structure"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1038\/s41586-019-1438-2","article-title":"Structure and mechanism of the cation-chloride cotransporter NKCC1","volume":"572","author":"Chew","year":"2019","journal-title":"Nature"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1038\/s41467-020-14790-3","article-title":"Structure of the human cation-chloride cotransporter NKCC1 determined by single-particle electron cryo-microscopy","volume":"11","author":"Yang","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1038\/s42003-021-01750-w","article-title":"The structural basis of function and regulation of neuronal cotransporters NKCC1 and KCC2","volume":"4","author":"Zhang","year":"2021","journal-title":"Commun. Biol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"2747","DOI":"10.1038\/s41467-022-30407-3","article-title":"Structural basis for inhibition of the Cation-chloride cotransporter NKCC1 by the diuretic drug bumetanide","volume":"13","author":"Zhao","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1126\/science.aay3129","article-title":"Cryo-EM structures of the human cation-chloride cotransporter KCC1","volume":"366","author":"Liu","year":"2019","journal-title":"Science"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"e107294","DOI":"10.15252\/embj.2020107294","article-title":"Phospho-regulation, nucleotide binding and ion access control in potassium-chloride cotransporters","volume":"40","author":"Chi","year":"2021","journal-title":"EMBO J."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"e2109083119","DOI":"10.1073\/pnas.2109083119","article-title":"Structure of the human cation-chloride cotransport KCC1 in an outward-open state","volume":"119","author":"Zhao","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1038\/s41422-021-00536-3","article-title":"Cryo-EM structures of the full-length human KCC2 and KCC3 cation-chloride cotransporters","volume":"31","author":"Chi","year":"2021","journal-title":"Cell Res."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"eabc5883","DOI":"10.1126\/sciadv.abc5883","article-title":"Structures and an activation mechanism of human potassium-chloride cotransporters","volume":"6","author":"Xie","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"e52505","DOI":"10.7554\/eLife.52505","article-title":"Cryo-EM structure of the potassium-chloride cotransporter KCC4 in lipid nanodiscs","volume":"9","author":"Reid","year":"2020","journal-title":"Elife"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"eadd7176","DOI":"10.1126\/sciadv.add7176","article-title":"Cryo-EM structure of the human sodium-chloride cotransporter NCC","volume":"8","author":"Nan","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"eabq0952","DOI":"10.1126\/sciadv.abq0952","article-title":"Inhibition mechanism of NKCC1 involves the carboxyl terminus and long-range conformational coupling","volume":"8","author":"Moseng","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_84","doi-asserted-by":"crossref","unstructured":"Zhao, Y., Shen, J., Wang, Q., Zhou, M., and Cao, E. (2020). Inhibitory and Transport Mechanisms of the Human Cation-Chloride Cotransport KCC1. bioRxiv, 221770.","DOI":"10.1101\/2020.07.26.221770"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"3718","DOI":"10.1021\/bi992301v","article-title":"The structural unit of the secretory Na+-K+-2Cl(-) cotransporter (NKCCl) is a homodimer","volume":"39","author":"Turner","year":"2000","journal-title":"Biochemistry"},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"18083","DOI":"10.1074\/jbc.M607811200","article-title":"Homooligomeric and heterooligomeric associations between K+-Cl- cotransporter isoforms and between K+-Cl- and Na+-K+-Cl- cotransporters","volume":"282","author":"Simard","year":"2007","journal-title":"J Biol. Chem."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"2210","DOI":"10.1074\/jbc.M111.309211","article-title":"Regulatory activation is accompanied by movements in the C-terminus of the Na-K-Cl cotransporter (NKCC1)","volume":"287","author":"Monette","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_88","doi-asserted-by":"crossref","unstructured":"Neumann, C., Rosenbaek, L.L., Flygaard, R.K., Habeck, M., Karlsen, J.L., Wang, Y., Lindorff-Larsen, K., Gad, H.H., Hartmann, R., and Lyons, J.A. (2022). Cryo-EM structure of the human NKCC1 transporter reveals mechanisms of ion coupling and specificity. Embo. J., e110169.","DOI":"10.1101\/2021.11.11.468215"},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1038\/nature08143","article-title":"Unlocking the molecular secrets of sodium-coupled transporters","volume":"459","author":"Krishnamurthy","year":"2009","journal-title":"Nature"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1126\/science.1160406","article-title":"The crystal structure of a sodium galactose transporter reveals mechanistic insights into Na(+)\/sugar symport","volume":"321","author":"Faham","year":"2008","journal-title":"Science"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1126\/science.1164440","article-title":"Structure and Molecular Mechanism of a Nucleobase-Cation-Symport-1 Family Transporter","volume":"322","author":"Weyand","year":"2008","journal-title":"Science"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.cell.2018.06.005","article-title":"Reactivation of Dormant Relay Pathways in Injured Spinal Cord by KCC2 Manipulations","volume":"174","author":"Chen","year":"2018","journal-title":"Cell"},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"1394","DOI":"10.1038\/nm.4442","article-title":"The small molecule CLP257 does not modify activity of the K+-Cl- co-transporter KCC2 but does potentiate GABA(A) receptor activity","volume":"23","author":"Cardarelli","year":"2017","journal-title":"Nat. Med."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"8434","DOI":"10.1016\/S0021-9258(19)83931-5","article-title":"[H-3] Bumetanide Binding to Duck Red-Cells-Correlation with Inhibition of (Na + K + 2cl) Cotransport","volume":"261","author":"Haas","year":"1986","journal-title":"J. Biol. Chem."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1016\/j.yebeh.2016.03.021","article-title":"The search for NKCC1-selective drugs for the treatment of epilepsy: Structure-function relationship of bumetanide and various bumetanide derivatives in inhibiting the human cation-chloride cotransporter NKCC1A","volume":"59","author":"Lykke","year":"2016","journal-title":"Epilepsy Behav."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.sbi.2019.03.011","article-title":"Insights into the mechanism and pharmacology of neurotransmitter sodium symporters","volume":"54","author":"Navratna","year":"2019","journal-title":"Curr. Opin. Struc. Biol."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1038\/nsmb.1854","article-title":"Ion\/substrate-dependent conformational dynamics of a bacterial homolog of neurotransmitter: Sodium symporters","volume":"17","author":"Claxton","year":"2010","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"863","DOI":"10.1113\/jphysiol.2013.259051","article-title":"How LeuT shapes our understanding of the mechanisms of sodium-coupled neurotransmitter transporters","volume":"592","author":"Penmatsa","year":"2014","journal-title":"J. Physiol.-Lond."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"U469","DOI":"10.1038\/nature10737","article-title":"X-ray structures of LeuT in substrate-free outward-open and apo inward-open states","volume":"481","author":"Krishnamurthy","year":"2012","journal-title":"Nature"},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1038\/s41586-019-1135-1","article-title":"Serotonin transporter-ibogaine complexes illuminate mechanisms of inhibition and transport","volume":"569","author":"Coleman","year":"2019","journal-title":"Nature"},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"17308","DOI":"10.1074\/jbc.M112.356014","article-title":"Loop Diuretic and Ion-binding Residues Revealed by Scanning Mutagenesis of Transmembrane Helix 3 (TM3) of Na-K-Cl Cotransporter (NKCC1)","volume":"287","author":"Somasekharan","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/S0014-5793(04)00328-X","article-title":"Interaction of neuron-specific K+-Cl- cotransporter, KCC2, with brain-type creatine kinase","volume":"564","author":"Inoue","year":"2004","journal-title":"FEBS Lett."},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1111\/j.1471-4159.2005.03560.x","article-title":"Brain-type creatine kinase activates neuron-specific K+-Cl- co-transporter KCC2","volume":"96","author":"Inoue","year":"2006","journal-title":"J. Neurochem."},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"3561","DOI":"10.1073\/pnas.1212907110","article-title":"Neto2 is a KCC2 interacting protein required for neuronal Cl- regulation in hippocampal neurons","volume":"110","author":"Ivakine","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1523\/JNEUROSCI.2926-20.2021","article-title":"Gephyrin Interacts with the K-Cl Cotransporter KCC2 to Regulate Its Surface Expression and Function in Cortical Neurons","volume":"42","author":"Donneger","year":"2022","journal-title":"J. Neurosci."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"re3","DOI":"10.1126\/scisignal.2005365","article-title":"The WNK-SPAK\/OSR1 pathway: Master regulator of cation-chloride cotransporters","volume":"7","author":"Alessi","year":"2014","journal-title":"Sci. Signal"},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"14115","DOI":"10.1074\/jbc.M110.112672","article-title":"Multiple pathways for protein phosphatase 1 (PP1) regulation of Na-K-2Cl cotransporter (NKCC1) function: The N-terminal tail of the Na-K-2Cl cotransporter serves as a regulatory scaffold for Ste20-related proline\/alanine-rich kinase (SPAK) AND PP1","volume":"285","author":"Gagnon","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"525","DOI":"10.1016\/j.cell.2009.05.031","article-title":"Sites of Regulated Phosphorylation that Control K-Cl Cotransporter Activity","volume":"138","author":"Rinehart","year":"2009","journal-title":"Cell"},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"37542","DOI":"10.1074\/jbc.M206293200","article-title":"A regulatory locus of phosphorylation in the N terminus of the Na-K-Cl cotransporter, NKCC1","volume":"277","author":"Darman","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"28755","DOI":"10.1074\/jbc.M603773200","article-title":"The Na+: Cl- cotransporter is activated and phosphorylated at the amino-terminal domain upon intracellular chloride depletion","volume":"281","author":"Cristobal","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"18668","DOI":"10.1074\/jbc.M114.567834","article-title":"A novel regulatory locus of phosphorylation in the C terminus of the potassium chloride cotransporter KCC2 that interferes with N-ethylmaleimide or staurosporine-mediated activation","volume":"289","author":"Weber","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_112","doi-asserted-by":"crossref","first-page":"16984","DOI":"10.1074\/jbc.RA118.004349","article-title":"Phosphoregulation of the intracellular termini of K(+)-Cl(-) cotransporter 2 (KCC2) enables flexible control of its activity","volume":"293","author":"Cordshagen","year":"2018","journal-title":"J. Biol. Chem."}],"container-title":["Membranes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0375\/12\/12\/1206\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:29:30Z","timestamp":1760146170000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0375\/12\/12\/1206"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,29]]},"references-count":112,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["membranes12121206"],"URL":"https:\/\/doi.org\/10.3390\/membranes12121206","relation":{},"ISSN":["2077-0375"],"issn-type":[{"value":"2077-0375","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,29]]}}}