{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T03:51:55Z","timestamp":1790999515955,"version":"4.1.0"},"reference-count":60,"publisher":"Microbiology Society","issue":"3","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,3,1]]},"abstract":"<jats:p>\n                    The p7 viroporin of the hepatitis C virus (HCV) forms an intracellular proton-conducting transmembrane channel in virus-infected cells, shunting the pH of intracellular compartments and thus helping virus assembly and release. This activity is essential for virus infectivity, making viroporins an attractive target for drug development. The protein sequence and drug sensitivity of p7 vary between the seven major genotypes of the hepatitis C virus, but the essential channel activity is preserved. Here, we investigated the effect of several inhibitors on recombinant HCV p7 channels corresponding to genotypes 1a\u2013b, 2a\u2013b, 3a and 4a using patch-clamp electrophysiology and cell-based assays. We established a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based cell viability assay for recombinant p7 expressed in HEK293 cells to assess channel activity and its sensitivity to inhibitors. The results from the cell viability assay were consistent with control measurements using established assays of haemadsorption and intracellular pH, and agreed with data from patch-clamp electrophysiology. Hexamethylene amiloride (HMA) was the most potent inhibitor of p7 activity, but possessed cytotoxic activity at higher concentrations. Rimantadine was active against p7 of all genotypes, while amantadine activity was genotype-dependent. The alkyl-chain iminosugars\n                    <jats:italic>N<\/jats:italic>\n                    B-DNJ,\n                    <jats:italic>N<\/jats:italic>\n                    N-DNJ and\n                    <jats:italic>N<\/jats:italic>\n                    N-DGJ were tested and their activity was found to be genotype-specific. In the current study, we introduce cell viability assays as a rapid and cost-efficient technique to assess viroporin activity and identify channel inhibitors as potential novel antiviral drugs.\n                  <\/jats:p>","DOI":"10.1099\/jgv.0.001571","type":"journal-article","created":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T10:17:02Z","timestamp":1615544222000},"source":"Crossref","is-referenced-by-count":14,"title":["Cell viability assay as a tool to study activity and inhibition of hepatitis C p7 channels"],"prefix":"10.1099","volume":"102","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3057-5919","authenticated-orcid":false,"given":"Ulrike","family":"Breitinger","sequence":"first","affiliation":[{"name":"Department of Biochemistry, German University in Cairo, New Cairo, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7759-4105","authenticated-orcid":false,"given":"Noha S.","family":"Farag","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, German University in Cairo, New Cairo, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1051-2558","authenticated-orcid":false,"given":"Nourhan K. M.","family":"Ali","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, German University in Cairo, New Cairo, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marwa","family":"Ahmed","sequence":"additional","affiliation":[{"name":"Present address: Institute of Biochemistry and Biophysics Friedrich-Schiller-University Jena, Hans-Kn\u00f6ll-Str. 2, D-07745, Jena, Germany"},{"name":"Department of Biochemistry, German University in Cairo, New Cairo, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed A.","family":"El-Azizi","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, German University in Cairo, New Cairo, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0111-3528","authenticated-orcid":false,"given":"Hans-Georg","family":"Breitinger","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, German University in Cairo, New Cairo, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"345","reference":[{"key":"R1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.27259"},{"key":"R2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2334-13-288"},{"key":"R3","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-8278(99)80374-2"},{"key":"R4","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v13.i17.2406"},{"key":"R5","doi-asserted-by":"publisher","DOI":"10.1007\/s00535-007-2030-3"},{"key":"R6","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.02831-14"},{"key":"R7","doi-asserted-by":"publisher","DOI":"10.3390\/v3112238"},{"key":"R8","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(03)00780-4"},{"key":"R9","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.76.8.3720-3730.2002"},{"key":"R10","doi-asserted-by":"publisher","DOI":"10.1002\/hep.22555"},{"key":"R11","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0030103"},{"key":"R12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1001087"},{"key":"R13","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(02)03851-6"},{"key":"R14","doi-asserted-by":"publisher","DOI":"10.1007\/0-387-25515-X_2"},{"key":"R15","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1031527100"},{"key":"R16","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(03)01453-4"},{"key":"R17","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2015.10.004"},{"key":"R18","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2007.05.001"},{"key":"R19","doi-asserted-by":"publisher","DOI":"10.1038\/nature12283"},{"key":"R20","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.122895"},{"key":"R21","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2016.04.018"},{"key":"R22","doi-asserted-by":"publisher","DOI":"10.1007\/s00249-009-0533-y"},{"key":"R23","doi-asserted-by":"publisher","DOI":"10.1021\/jm050721e"},{"key":"R24","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2015.03.006"},{"key":"R25","doi-asserted-by":"publisher","DOI":"10.1007\/s10858-015-9912-0"},{"key":"R26","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-019-44413-x"},{"key":"R27","doi-asserted-by":"publisher","DOI":"10.1111\/j.1572-0241.2005.00253.x"},{"key":"R28","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2005.11.038"},{"key":"R29","doi-asserted-by":"publisher","DOI":"10.1002\/hep.27298"},{"key":"R30","doi-asserted-by":"publisher","DOI":"10.1002\/hep.24171"},{"key":"R31","doi-asserted-by":"publisher","DOI":"10.1038\/ctg.2016.50"},{"key":"R32","doi-asserted-by":"publisher","DOI":"10.4155\/fmc.14.121"},{"key":"R33","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.2002.028910"},{"key":"R34","first-page":"712","article-title":"The p7 viroporin of the hepatitis C virus contributes to liver inflammation by stimulating production of interleukin-1\u03b2","volume":"2017","author":"Farag","year":"1863","journal-title":"Biochim Biophys Acta Mol Basis Dis"},{"key":"R35","doi-asserted-by":"publisher","DOI":"10.1099\/vir.0.19634-0"},{"key":"R36","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot095505"},{"key":"R37","doi-asserted-by":"publisher","DOI":"10.1177\/108705719900400206"},{"key":"R38","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(02)03851-6"},{"key":"R39","doi-asserted-by":"publisher","DOI":"10.1113\/jphysiol.1996.sp021495"},{"key":"R40","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1994.1628"},{"key":"R41","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(95)80003-2"},{"key":"R42","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.67.9.5585-5594.1993"},{"key":"R43","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201507112"},{"key":"R44","doi-asserted-by":"publisher","DOI":"10.1038\/srep08576"},{"key":"R45","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.68.2.920-926.1994"},{"key":"R46","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.68.2.911-919.1994"},{"key":"R47","doi-asserted-by":"publisher","DOI":"10.1002\/hep.21686"},{"key":"R48","doi-asserted-by":"publisher","DOI":"10.1006\/viro.2001.1121"},{"key":"R49","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.03693-14"},{"key":"R50","doi-asserted-by":"publisher","DOI":"10.5582\/bst.2014.01140"},{"key":"R51","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2015.03.036"},{"key":"R52","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.75.16.7481-7488.2001"},{"key":"R53","doi-asserted-by":"publisher","DOI":"10.1002\/hep.24371"},{"key":"R54","doi-asserted-by":"publisher","DOI":"10.1002\/hep.26685"},{"key":"R55","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1177\/135965350601100408","article-title":"Amino acid variations in hepatitis C virus p7 and sensitivity to antiviral combination therapy with amantadine in chronic hepatitis C","volume":"11","author":"Mihm","year":"2006","journal-title":"Antivir Ther"},{"key":"R56","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2036.2006.02863.x"},{"key":"R57","doi-asserted-by":"publisher","DOI":"10.1007\/s13238-016-0256-7"},{"key":"R58","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.662452"},{"key":"R59","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-381290-2.00004-5"},{"key":"R60","doi-asserted-by":"publisher","DOI":"10.1007\/978-981-10-8727-1_20"}],"container-title":["Journal of General Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jgv\/10.1099\/jgv.0.001571?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T04:06:58Z","timestamp":1671595618000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jgv\/10.1099\/jgv.0.001571"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,1]]},"references-count":60,"journal-issue":{"issue":"3","published-print":{"date-parts":[[2021,3,1]]}},"URL":"https:\/\/doi.org\/10.1099\/jgv.0.001571","relation":{},"ISSN":["0022-1317","1465-2099"],"issn-type":[{"value":"0022-1317","type":"print"},{"value":"1465-2099","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,1]]}}}