{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T22:07:52Z","timestamp":1780956472021,"version":"3.54.1"},"reference-count":95,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2022,9,27]],"date-time":"2022-09-27T00:00:00Z","timestamp":1664236800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"SEP-Cinvestav","award":["FIDSC 2018\/95"],"award-info":[{"award-number":["FIDSC 2018\/95"]}]},{"name":"SEP\u2013CONACYT","award":["A1-S-39422"],"award-info":[{"award-number":["A1-S-39422"]}]},{"DOI":"10.13039\/501100013704","name":"Consejo Nacional de Ciencia y Tecnolog\u00eda, Paraguay","doi-asserted-by":"publisher","award":["Grant 282663"],"award-info":[{"award-number":["Grant 282663"]}],"id":[{"id":"10.13039\/501100013704","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["2018-04165"],"award-info":[{"award-number":["2018-04165"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p><jats:italic>Giardia duodenalis<\/jats:italic>causes giardiasis, a major diarrheal disease in humans worldwide whose treatment relies mainly on metronidazole (MTZ) and albendazole (ABZ). The emergence of ABZ resistance in this parasite has prompted studies to elucidate the molecular mechanisms underlying this phenomenon.<jats:italic>G<\/jats:italic>.<jats:italic>duodenalis<\/jats:italic>trophozoites convert ABZ into its sulfoxide (ABZSO) and sulfone (ABZSOO) forms, despite lacking canonical enzymes involved in these processes, such as cytochrome P450s (CYP450s) and flavin-containing monooxygenases (FMOs). This study aims to identify the enzyme responsible for ABZ metabolism and its role in ABZ resistance in<jats:italic>G<\/jats:italic>.<jats:italic>duodenalis<\/jats:italic>. We first determined that the iron-containing cofactor heme induces higher mRNA expression levels of flavohemoglobin (gFlHb) in<jats:italic>Giardia<\/jats:italic>trophozoites. Molecular docking analyses predict favorable interactions of gFlHb with ABZ, ABZSO and ABZSOO. Spectral analyses of recombinant gFlHb in the presence of ABZ, ABZSO and ABZSOO showed high affinities for each of these compounds with<jats:italic>K<\/jats:italic><jats:sub><jats:italic>d<\/jats:italic><\/jats:sub>values of 22.7, 19.1 and 23.8 nM respectively. ABZ and ABZSO enhanced gFlHb NADH oxidase activity (turnover number 14.5 min<jats:sup>-1<\/jats:sup>), whereas LC-MS\/MS analyses of the reaction products showed that gFlHb slowly oxygenates ABZ into ABZSO at a much lower rate (turnover number 0.01 min<jats:sup>-1<\/jats:sup>). Further spectroscopic analyses showed that ABZ is indirectly oxidized to ABZSO by superoxide generated from the NADH oxidase activity of gFlHb. In a similar manner, the superoxide-generating enzyme xanthine oxidase was able to produce ABZSO in the presence of xanthine and ABZ. Interestingly, we find that gFlHb mRNA expression is lower in albendazole-resistant clones compared to those that are sensitive to this drug. Furthermore, all albendazole-resistant clones transfected to overexpress gFlHb displayed higher susceptibility to the drug than the parent clones. Collectively these findings indicate a role for gFlHb in ABZ conversion to its sulfoxide and that gFlHb down-regulation acts as a passive pharmacokinetic mechanism of resistance in this parasite.<\/jats:p>","DOI":"10.1371\/journal.ppat.1010840","type":"journal-article","created":{"date-parts":[[2022,9,27]],"date-time":"2022-09-27T17:36:36Z","timestamp":1664300196000},"page":"e1010840","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":11,"title":["Giardia duodenalis: Flavohemoglobin is involved in drug biotransformation and resistance to albendazole"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1579-2309","authenticated-orcid":true,"given":"Edar O.","family":"Pech-Santiago","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ra\u00fal","family":"Arg\u00fcello-Garc\u00eda","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7783-1708","authenticated-orcid":true,"given":"Citlali","family":"V\u00e1zquez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Saavedra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9229-1121","authenticated-orcid":true,"given":"Iliana","family":"Gonz\u00e1lez-Hern\u00e1ndez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2032-4753","authenticated-orcid":true,"given":"Helgi","family":"Jung-Cook","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6525-7435","authenticated-orcid":true,"given":"Steven P.","family":"Rafferty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1997-3211","authenticated-orcid":true,"given":"M. Guadalupe","family":"Ortega-Pierres","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2022,9,27]]},"reference":[{"key":"ppat.1010840.ref001","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1038\/nrmicro2317","article-title":"Behind the smile: cell biology and disease mechanisms of Giardia species","volume":"8","author":"J Ankarklev","year":"2010","journal-title":"Nat Rev Microbiol"},{"key":"ppat.1010840.ref002","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1016\/j.pt.2009.11.010","article-title":"Giardiasis\u2014why do the symptoms sometimes never stop?","volume":"26","author":"LJ Robertson","year":"2010","journal-title":"Trends Parasitol"},{"key":"ppat.1010840.ref003","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.pt.2006.02.015","article-title":"Giardia and Cryptosporidium join the \u201cNeglected Diseases Initiative.\u201d","volume":"22","author":"L Savioli","year":"2006","journal-title":"Trends Parasitol"},{"key":"ppat.1010840.ref004","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/j.ijpddr.2015.03.001","article-title":"Comparative characterisation of two nitroreductases from Giardia lamblia as potential activators of nitro compounds","volume":"5","author":"J M\u00fcller","year":"2015","journal-title":"Int J Parasitol Drugs Drug Resist"},{"key":"ppat.1010840.ref005","first-page":"841","article-title":"Giardiasis: Recent progress in chemotherapy and drug development","volume":"9","author":"T Mineno","year":"2003","journal-title":"Curr Pharm Des"},{"key":"ppat.1010840.ref006","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/j.molbiopara.2004.08.001","article-title":"Characterisation of benzimidazole binding with recombinant tubulin from Giardia duodenalis, Encephalitozoon intestinalis, and Cryptosporidium parvum","volume":"138","author":"LM MacDonald","year":"2004","journal-title":"Mol Biochem Parasitol"},{"key":"ppat.1010840.ref007","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.jmgm.2004.08.001","article-title":"A possible model of benzimidazole binding to \u03b2-tubulin disclosed by invoking an inter-domain movement","volume":"23","author":"MW Robinson","year":"2004","journal-title":"J Mol Graph Model"},{"key":"ppat.1010840.ref008","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1007\/s00228-003-0636-9","article-title":"Identification of human cytochrome P450s that metabolise anti-parasitic drugs and predictions of in vivo drug hepatic clearance from in vitro data","volume":"59","author":"XQ Li","year":"2003","journal-title":"Eur J Clin Pharmacol"},{"key":"ppat.1010840.ref009","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1046\/j.1365-2125.2000.00170.x","article-title":"Relative contribution of cytochromes P 450 and flavin containing monoxygenases to the metabolism of albendazole by human liver microsomes","volume":"49","author":"HC Rawden","year":"2000","journal-title":"Br J Clin Pharmacol"},{"key":"ppat.1010840.ref010","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1007\/978-3-319-12108-6_2","volume-title":"Electron Transfer Partners of Cytochrome P450 BT\u2014Cytochrome P450: Structure, Mechanism, and Biochemistry","author":"L Waskell","year":"2015"},{"key":"ppat.1010840.ref011","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fmicb.2015.00286","article-title":"An antioxidant response is involved in resistance of Giardia duodenalis to albendazole","volume":"6","author":"R Arg\u00fcello-Garc\u00eda","year":"2015","journal-title":"Front Microbiol"},{"key":"ppat.1010840.ref012","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1371\/journal.pone.0017285","article-title":"The minimal proteome in the reduced mitochondrion of the parasitic protist Giardia intestinalis","volume":"6","author":"PL Jedelsk\u00fd","year":"2011","journal-title":"PLoS One"},{"key":"ppat.1010840.ref013","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1039\/c2mt20152f","article-title":"Cytochrome b5 from Giardia lamblia","volume":"4","author":"S Alam","year":"2012","journal-title":"Metallomics"},{"key":"ppat.1010840.ref014","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1128\/EC.00200-13","article-title":"Giardia intestinalis incorporates heme into cytosolic cytochrome b5","volume":"13","author":"J Pyrih","year":"2014","journal-title":"Eukaryot Cell"},{"key":"ppat.1010840.ref015","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1016\/0035-9203(93)90296-3","article-title":"Albendazole is highly effective against cutaneous larva migrans but not against Giardia infection: results of an open pilot trial in travellers returning from the tropics","volume":"87","author":"H Kollaritsch","year":"1993","journal-title":"Trans R Soc Trop Med Hyg"},{"key":"ppat.1010840.ref016","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1086\/320886","article-title":"Treatment of patients with refractory giardiasis","volume":"33","author":"TE Nash","year":"2001","journal-title":"Clin Infect Dis"},{"key":"ppat.1010840.ref017","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1159\/000104728","article-title":"Therapy-resistant diarrhea due to Giardia lamblia in a patient with common variable immunodeficiency disease","volume":"75","author":"M Sawatzki","year":"2007","journal-title":"Digestion"},{"key":"ppat.1010840.ref018","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1016\/j.meegid.2009.05.015","article-title":"In vitro resistance to 5-nitroimidazoles and benzimidazoles in Giardia duodenalis: Variability and variation in gene expression","volume":"9","author":"R Arg\u00fcello-Garc\u00eda","year":"2009","journal-title":"Infect Genet Evol"},{"key":"ppat.1010840.ref019","first-page":"57","article-title":"Persistence of intestinal parasitic infections during the national de-worming campaign in schoolchildren of northwestern Mexico: A cross-sectional study","volume":"25","author":"L Quihui-Cota","year":"2012","journal-title":"Ann Gastroenterol"},{"key":"ppat.1010840.ref020","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/0166-6851(90)90025-H","article-title":"Specific interaction of benzimidazole anthelmintics with tubulin: high-affinity binding and benzimidazole resistance in Haemonchus contortus","volume":"38","author":"GW Lubega","year":"1990","journal-title":"Mol Biochem Parasitol"},{"key":"ppat.1010840.ref021","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pntd.0002247","article-title":"Association between Response to Albendazole Treatment and B-Tubulin Genotype Frequencies in Soil-transmitted Helminths","volume":"7","author":"A Diawara","year":"2013","journal-title":"PLoS Negl Trop Dis"},{"key":"ppat.1010840.ref022","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1186\/1756-3305-6-236","article-title":"Genetic variations in the beta-tubulin gene and the internal transcribed spacer 2 region of Trichuris species from man and baboons","volume":"6","author":"TV Hansen","year":"2013","journal-title":"Parasit Vectors"},{"key":"ppat.1010840.ref023","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1016\/j.ijpara.2015.04.007","article-title":"Genetic evidence for the spread of a benzimidazole resistance mutation across southern India from a single origin in the parasitic nematode Haemonchus contortus","volume":"45","author":"U Chaudhry","year":"2015","journal-title":"Int J Parasitol"},{"key":"ppat.1010840.ref024","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1089\/mdr.1996.2.303","article-title":"Albendazole resistance in Giardia is correlated with cytoskeletal changes but not with a mutation at amino acid 200 in beta-tubulin","volume":"2","author":"Upcroft","year":"1996","journal-title":"Microb Drug Resist"},{"key":"ppat.1010840.ref025","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.ijpddr.2021.05.003","article-title":"In vitro selection of Giardia duodenalis for Albendazole resistance identifies a \u03b2-tubulin mutation at amino acid E198K","volume":"16","author":"SJ Emery-Corbin","year":"2021","journal-title":"Int J Parasitol Drugs Drug Resist"},{"key":"ppat.1010840.ref026","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.meegid.2012.08.021","article-title":"Proteomic and transcriptional analyses of genes differentially expressed in Giardia duodenalis clones resistant to albendazole","volume":"15","author":"MT Paz-Maldonado","year":"2013","journal-title":"Infect Genet Evol"},{"key":"ppat.1010840.ref027","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3389\/fmicb.2015.00800","article-title":"Albendazole induces oxidative stress and DNA damage in the parasitic protozoan Giardia duodenalis","volume":"6","author":"R Mart\u00ednez-Espinosa","year":"2015","journal-title":"Front Microbiol"},{"key":"ppat.1010840.ref028","doi-asserted-by":"crossref","first-page":"4643","DOI":"10.1128\/AAC.00384-11","article-title":"Impaired parasite attachment as fitness cost of metronidazole resistance in Giardia lamblia","volume":"55","author":"N Tejman-Yarden","year":"2011","journal-title":"Antimicrob Agents Chemother"},{"issue":"80-","key":"ppat.1010840.ref029","doi-asserted-by":"crossref","first-page":"1921","DOI":"10.1126\/science.1143837","article-title":"Genomic minimalism in the early diverging intestinal parasite Giardia lamblia","volume":"317","author":"HG Morrison","year":"2007","journal-title":"Science"},{"key":"ppat.1010840.ref030","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1016\/j.bbrc.2010.07.073","article-title":"Giardia lamblia encodes a functional flavohemoglobin","volume":"399","author":"S Rafferty","year":"2010","journal-title":"Biochem Biophys Res Commun"},{"key":"ppat.1010840.ref031","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.jinorgbio.2004.10.003","article-title":"Nitric oxide dioxygenase function and mechanism of flavohemoglobin, hemoglobin, myoglobin and their associated reductases","volume":"99","author":"PR Gardner","year":"2005","journal-title":"J Inorg Biochem"},{"key":"ppat.1010840.ref032","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1471-2148-6-31","article-title":"A phylogenomic profile of globins","volume":"6","author":"SN Vinogradov","year":"2006","journal-title":"BMC Evol Biol"},{"key":"ppat.1010840.ref033","doi-asserted-by":"crossref","first-page":"654","DOI":"10.1016\/j.bbrc.2010.07.137","article-title":"Flavohemoglobin and nitric oxide detoxification in the human protozoan parasite Giardia intestinalis","volume":"399","author":"D Mastronicola","year":"2010","journal-title":"Biochem Biophys Res Commun"},{"key":"ppat.1010840.ref034","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1021\/bi101650q","article-title":"Structure of Ralstonia eutropha flavohemoglobin in complex with three antibiotic azole compounds","volume":"50","author":"E El Hammi","year":"2011","journal-title":"Biochemistry"},{"key":"ppat.1010840.ref035","doi-asserted-by":"crossref","first-page":"16435","DOI":"10.1074\/jbc.M111.329920","article-title":"An unconventional hexacoordinated flavohemoglobin from Mycobacterium tuberculosis","volume":"287","author":"S Gupta","year":"2012","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref036","doi-asserted-by":"crossref","first-page":"22272","DOI":"10.1074\/jbc.M301285200","article-title":"Escherichia coli flavohemoglobin is an efficient alkylhydroperoxide reductase","volume":"278","author":"A Bonamore","year":"2003","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref037","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1016\/S0014-5793(99)00157-X","article-title":"Anoxic function for the Escherichia coli flavohaemoglobin (Hmp): Reversible binding of nitric oxide and reduction to nitrous oxide.","volume":"445","author":"SO Kim","year":"1999","journal-title":"FEBS Lett"},{"key":"ppat.1010840.ref038","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1093\/nar\/gkn631","article-title":"GiardiaDB and TrichDB: Integrated genomic resources for the eukaryotic protist pathogens Giardia lamblia and Trichomonas vaginalis","volume":"37","author":"C Aurrecoechea","year":"2009","journal-title":"Nucleic Acids Res"},{"key":"ppat.1010840.ref039","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1002\/pro.3290","article-title":"Clustal Omega for making accurate alignments of many protein sequences","volume":"27","author":"F Sievers","year":"2017","journal-title":"Protein Sci"},{"key":"ppat.1010840.ref040","doi-asserted-by":"crossref","first-page":"W174","DOI":"10.1093\/nar\/gkv342","article-title":"I-TASSER server: New development for protein structure and function predictions","volume":"43","author":"J Yang","year":"2015","journal-title":"Nucleic Acids Res"},{"key":"ppat.1010840.ref041","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1016\/S0076-6879(97)77022-8","article-title":"VERIFY3D: Assessment of protein models with three-dimensional profiles","volume":"277","author":"D Eisenberg","year":"1997","journal-title":"Methods Enzymol"},{"issue":"80-","key":"ppat.1010840.ref042","doi-asserted-by":"crossref","first-page":"164 LP","DOI":"10.1126\/science.1853201","article-title":"A method to identify protein sequences that fold into a known three-dimensional structure","volume":"253","author":"JU Bowie","year":"1991","journal-title":"Science"},{"key":"ppat.1010840.ref043","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1038\/356083a0","article-title":"Assessment of protein models with three-dimensional profiles","volume":"356","author":"R L\u00fcthy","year":"1992","journal-title":"Nature"},{"key":"ppat.1010840.ref044","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1093\/nar\/gkm290","article-title":"ProSA-web: Interactive web service for the recognition of errors in three-dimensional structures of proteins","volume":"35","author":"M Wiederstein","year":"2007","journal-title":"Nucleic Acids Res"},{"key":"ppat.1010840.ref045","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1006\/jmbi.1999.3091","article-title":"Protein secondary structure prediction based on position-specific scoring matrices11Edited by G","volume":"292","author":"DT Jones","year":"1999","journal-title":"Von Heijne. J Mol Biol"},{"key":"ppat.1010840.ref046","doi-asserted-by":"crossref","first-page":"2302","DOI":"10.1093\/nar\/gki524","article-title":"TM-align: A protein structure alignment algorithm based on the TM-score","volume":"33","author":"Y Zhang","year":"2005","journal-title":"Nucleic Acids Res"},{"key":"ppat.1010840.ref047","doi-asserted-by":"crossref","first-page":"2324","DOI":"10.1021\/acs.jcim.5b00559","article-title":"ZINC 15 \u2013Ligand Discovery for Everyone","volume":"55","author":"T Sterling","year":"2015","journal-title":"J Chem Inf Model"},{"key":"ppat.1010840.ref048","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1093\/nar\/gkr366","article-title":"SwissDock, a protein-small molecule docking web service based on EADock DSS","volume":"39","author":"A Grosdidier","year":"2011","journal-title":"Nucleic Acids Res"},{"key":"ppat.1010840.ref049","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1002\/jcc.21334","article-title":"AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading","volume":"31","author":"O Trott","year":"2010","journal-title":"J Comput Chem"},{"key":"ppat.1010840.ref050","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/0014-5793(96)00154-8","article-title":"The flavohaemoglobin (HMP) of Escherichia coli generates superoxide in vitro and causes oxidative stress in vivo","volume":"382","author":"J Membrillo-Hern\u00e1ndez","year":"1996","journal-title":"FEBS Lett"},{"key":"ppat.1010840.ref051","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1016\/0003-2697(71)90370-8","article-title":"Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels","volume":"44","author":"C Beauchamp","year":"1971","journal-title":"Anal Biochem"},{"key":"ppat.1010840.ref052","first-page":"267","article-title":"A simple LC\u2014MS \/ MS method to determine plasma and cerebrospinal fl uid levels of albendazole metabolites (albendazole sulfoxide and albendazole sulfone) in patients with neurocysticercosis","author":"I Gonz\u00e1lez-Hern\u00e1ndez","year":"2012"},{"key":"ppat.1010840.ref053","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/S0166-6851(97)00225-9","article-title":"Episomal and integrated maintenance of foreign DNA in Giardia lamblia","volume":"92","author":"SM Singer","year":"1998","journal-title":"Mol Biochem Parasitol"},{"key":"ppat.1010840.ref054","doi-asserted-by":"crossref","first-page":"4125","DOI":"10.1021\/bi047389d","article-title":"Ligand binding properties of bacterial hemoglobins and flavohemoglobins","volume":"44","author":"J Farr\u00e9s","year":"2005","journal-title":"Biochemistry"},{"key":"ppat.1010840.ref055","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.1128\/AAC.49.5.1837-1843.2005","article-title":"Imidazole antibiotics inhibit the nitric oxide dioxygenase function of microbial flavohemoglobin","volume":"49","author":"RA Helmick","year":"2005","journal-title":"Antimicrob Agents Chemother"},{"key":"ppat.1010840.ref056","doi-asserted-by":"crossref","first-page":"1527","DOI":"10.1128\/JB.01378-09","article-title":"Binding of azole antibiotics to Staphylococcus aureus flavohemoglobin increases intracellular oxidative stress","volume":"192","author":"LS Nobre","year":"2010","journal-title":"J Bacteriol"},{"key":"ppat.1010840.ref057","doi-asserted-by":"crossref","first-page":"4565","DOI":"10.1111\/febs.12043","article-title":"Active site analysis of yeast flavohemoglobin based on its structure with a small ligand or econazole","volume":"279","author":"E El Hammi","year":"2012","journal-title":"FEBS J"},{"key":"ppat.1010840.ref058","article-title":"Sequence-structure analysis of FAD-containing proteins","author":"O Dym","year":"2001","journal-title":"Protein Sci"},{"key":"ppat.1010840.ref059","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1038\/nmeth.3213","article-title":"The I-TASSER Suite: Protein structure and function prediction","volume":"12","author":"J Yang","year":"2015","journal-title":"Nat Methods"},{"key":"ppat.1010840.ref060","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1107\/S0021889892009944","article-title":"PROCHECK: a program to check the stereochemical quality of protein structures","volume":"26","author":"RA Laskowski","year":"1993","journal-title":"J Appl Crystallogr"},{"key":"ppat.1010840.ref061","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1093\/nar\/gkp322","article-title":"QMEAN server for protein model quality estimation","volume":"37","author":"P Benkert","year":"2009","journal-title":"Nucleic Acids Res"},{"key":"ppat.1010840.ref062","doi-asserted-by":"crossref","first-page":"7272","DOI":"10.1074\/jbc.M009280200","article-title":"Flavohemoglobin, a Globin with a Peroxidase-like Catalytic Site","volume":"276","author":"M Mukai","year":"2001","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref063","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1002\/iub.9","article-title":"Flavohemoglobin: Structure and reactivity","volume":"60","author":"A Bonamore","year":"2008","journal-title":"IUBMB Life"},{"key":"ppat.1010840.ref064","doi-asserted-by":"crossref","first-page":"295","DOI":"10.2174\/187152610791591629","article-title":"Giardiasis, drug resistance, and new target discovery","volume":"10","author":"HF Tian","year":"2010","journal-title":"Infect Disord Drug Targets"},{"key":"ppat.1010840.ref065","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1128\/CMR.14.1.150-164.2001","article-title":"Drug Targets and Mechanisms of Resistance in the Anaerobic Protozoa Drug Targets and Mechanisms of Resistance in the Anaerobic Protozoa","volume":"14","author":"P Upcroft","year":"2001","journal-title":"Clin Microbiol Rev"},{"key":"ppat.1010840.ref066","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1128\/AAC.50.1.344-347.2006","article-title":"5-Nitroimidazole drugs effective against metronidazole-resistant Trichomonas vaginalis and Giardia duodenalis","volume":"50","author":"JA Upcroft","year":"2006","journal-title":"Antimicrob Agents Chemother"},{"key":"ppat.1010840.ref067","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1017\/S0031182000065823","article-title":"Quinacrine-resistant Giardia duodenalis","volume":"112","author":"JA Upcroft","year":"1996","journal-title":"Parasitology"},{"key":"ppat.1010840.ref068","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1093\/jac\/dkm205","article-title":"Characterization of Giardia lamblia WB C6 clones resistant to nitazoxanide and to metronidazole","volume":"60","author":"J M\u00fcller","year":"2007","journal-title":"J Antimicrob Chemother"},{"key":"ppat.1010840.ref069","doi-asserted-by":"crossref","first-page":"1885","DOI":"10.1517\/14656566.8.12.1885","article-title":"Giardiasis: A pharmacotherapy review","volume":"8","author":"AA Escobedo","year":"2007","journal-title":"Expert Opin Pharmacother"},{"key":"ppat.1010840.ref070","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.2174\/1568026618666181002095314","article-title":"Giardiasis: Characteristics, Pathogenesis and New Insights About Treatment","volume":"18","author":"V Vivancos","year":"2018","journal-title":"Curr Top Med Chem"},{"key":"ppat.1010840.ref071","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1093\/ajcn\/73.1.53","article-title":"Anthelmintic treatment of rural bangladeshi children: Effect on host physiology, growth, and biochemical status","volume":"73","author":"CA Northrop-Clewes","year":"2001","journal-title":"Am J Clin Nutr"},{"key":"ppat.1010840.ref072","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1111\/j.1550-7408.1992.tb04841.x","article-title":"Giardia lamblia: Ultrastructural study of the In vitro Effect of Benzimidazoles","volume":"39","author":"B Chavez","year":"1992","journal-title":"J Protozol"},{"key":"ppat.1010840.ref073","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1046\/j.1365-2885.2000.00254.x","article-title":"The binding and distribution of albendazole and its principal metabolites in Giardia duodenalis","volume":"23","author":"ME Oxberry","year":"2000","journal-title":"J Vet Pharmacol Ther"},{"key":"ppat.1010840.ref074","first-page":"398","article-title":"Transcriptomics indicates active and passive metronidazole resistance mechanisms in three seminal Giardia lines","volume":"8","author":"BRE Ansell","year":"2017","journal-title":"Front Microbiol"},{"key":"ppat.1010840.ref075","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1128\/AAC.37.2.328","article-title":"Activities of several benzimidazoles and tubulin inhibitors against Giardia spp. in vitro","volume":"37","author":"UM Morgan","year":"1993","journal-title":"Antimicrob Agents Chemother"},{"key":"ppat.1010840.ref076","doi-asserted-by":"crossref","first-page":"1700","DOI":"10.1161\/01.ATV.11.6.1700","article-title":"Hemin: A possible physiological mediator of low density lipoprotein oxidation and endothelial injury","volume":"11","author":"G Balla","year":"1991","journal-title":"Arterioscler Thromb Vasc Biol"},{"key":"ppat.1010840.ref077","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1080\/019021400269871","article-title":"Hemoglobin potentiates the production of reactive oxygen species by alveolar macrophages","volume":"26","author":"LJ Huffman","year":"2000","journal-title":"Exp Lung Res"},{"key":"ppat.1010840.ref078","doi-asserted-by":"crossref","first-page":"804","DOI":"10.1016\/j.neuint.2011.03.006","article-title":"Uptake, metabolism and toxicity of hemin in cultured neurons","volume":"58","author":"TN Dang","year":"2011","journal-title":"Neurochem Int"},{"key":"ppat.1010840.ref079","doi-asserted-by":"crossref","first-page":"1540","DOI":"10.1002\/glia.21198","article-title":"The metabolism and toxicity of hemin in astrocytes","volume":"59","author":"TN Dang","year":"2011","journal-title":"Glia"},{"key":"ppat.1010840.ref080","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1002\/jnr.10633","article-title":"Hemin induces an iron-dependent, oxidative injury to human neuron-like cells","volume":"73","author":"L Goldstein","year":"2003","journal-title":"J Neurosci Res"},{"key":"ppat.1010840.ref081","doi-asserted-by":"crossref","first-page":"1394","DOI":"10.1152\/ajpheart.00584.2011","article-title":"Hemin causes mitochondrial dysfunction in endothelial cells through promoting lipid peroxidation: The protective role of autophagy","volume":"302","author":"AN Higdon","year":"2012","journal-title":"Am J Physiol\u2014Hear Circ Physiol"},{"key":"ppat.1010840.ref082","first-page":"880","article-title":"Formation of novel N-acetylcysteine-hemin adducts abrogates hemin-induced cytotoxicity and suppresses the NRF2-driven stress response in human pro-erythroid K562 cells","author":"SK Georgiou-Siafis","year":"2020","journal-title":"Eur J Pharmacol"},{"key":"ppat.1010840.ref083","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.exppara.2015.08.001","article-title":"Heme proteins of Giardia intestinalis","volume":"159","author":"SP Rafferty","year":"2015","journal-title":"Exp Parasitol"},{"key":"ppat.1010840.ref084","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1016\/j.bbrc.2004.03.007","article-title":"Mapping protein matrix cavities in human cytoglobin through Xe atom binding","volume":"316","author":"D de Sanctis","year":"2004","journal-title":"Biochem Biophys Res Commun"},{"key":"ppat.1010840.ref085","article-title":"Structural Studies on Flavohemoglobins. Methods in Enzymology","author":"A Ilari","year":"2008","journal-title":"Elsevier Masson SAS"},{"key":"ppat.1010840.ref086","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1007\/s00775-017-1487-7","article-title":"Resonance Raman studies on the flavohemoglobin of the protist Giardia intestinalis: evidence of a type I\/II-peroxidase-like heme environment and roles of the active site distal residues","volume":"22","author":"B Lukaszewicz","year":"2017","journal-title":"J Biol Inorg Chem"},{"key":"ppat.1010840.ref087","doi-asserted-by":"crossref","first-page":"10378","DOI":"10.1073\/pnas.95.18.10378","article-title":"Nitric oxide dioxygenase: An enzymic function for flavohemoglobin","volume":"95","author":"PR Gardner","year":"1998","journal-title":"Proc Natl Acad Sci U S A"},{"key":"ppat.1010840.ref088","first-page":"193","article-title":"Escherichia coli Hmp, an \u201coxygen-binding flavohaemoprotein\u201d, produces superoxide anion and self-destructs","volume":"182","author":"G Wu","year":"2004","journal-title":"Arch Microbiol"},{"key":"ppat.1010840.ref089","doi-asserted-by":"crossref","first-page":"100186","DOI":"10.1074\/jbc.RA120.016637","article-title":"Allostery in the nitric oxide dioxygenase mechanism of flavohemoglobin","volume":"296","author":"AM Gardner","year":"2021","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref090","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1002\/iub.1275","article-title":"Type i flavohemoglobin of mycobacterium smegmatis is a functional nitric oxide dioxygenase","volume":"66","author":"N Thakur","year":"2014","journal-title":"IUBMB Life"},{"key":"ppat.1010840.ref091","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.freeradbiomed.2004.04.031","article-title":"Nitric oxide metabolism in mammalian cells: Substrate and inhibitor profiles of a NADPH-cytochrome P450 oxidoreductase-coupled microsomal nitric oxide dioxygenase","volume":"37","author":"CK Hallstrom","year":"2004","journal-title":"Free Radic Biol Med"},{"key":"ppat.1010840.ref092","doi-asserted-by":"crossref","first-page":"6991","DOI":"10.1074\/jbc.270.12.6991","article-title":"Regulation of Saccharomyces cerevisiae flavohemoglobin gene expression","volume":"270","author":"MJ Crawford","year":"1995","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref093","doi-asserted-by":"crossref","first-page":"25131","DOI":"10.1074\/jbc.271.41.25131","article-title":"Function and Expression of Flavohemoglobin in Saccharomyces cerevisiae. Evidence for a role in the oxidative stress response","volume":"271","author":"XJ Zhao","year":"1996","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref094","doi-asserted-by":"crossref","first-page":"9527","DOI":"10.1074\/jbc.273.16.9527","article-title":"Flavohemoglobin Expression and Function in Saccharomyces cerevisiae","volume":"273","author":"N Buisson","year":"1998","journal-title":"J Biol Chem"},{"key":"ppat.1010840.ref095","first-page":"715","article-title":"Inducible Defense Mechanism against Nitric Oxide in Candida albicans Inducible Defense Mechanism against Nitric Oxide in Candida albicans \u2020","volume":"3","author":"BD Ullmann","year":"2004","journal-title":"Am J Infect Dis"}],"container-title":["PLOS Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1010840","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,27]],"date-time":"2023-11-27T20:52:15Z","timestamp":1701118335000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.ppat.1010840"}},"subtitle":[],"editor":[{"given":"Christine","family":"Clayton","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2022,9,27]]},"references-count":95,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2022,9,27]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.ppat.1010840","relation":{},"ISSN":["1553-7374"],"issn-type":[{"value":"1553-7374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,27]]}}}