{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T09:35:50Z","timestamp":1768556150033,"version":"3.49.0"},"reference-count":43,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2021,8,31]],"date-time":"2021-08-31T00:00:00Z","timestamp":1630368000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016928","name":"Instituto Nacional de Pediatria","doi-asserted-by":"publisher","award":["2019\/062"],"award-info":[{"award-number":["2019\/062"]}],"id":[{"id":"10.13039\/100016928","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Giardiasis represents a latent problem in public health due to the exceptionally pathogenic strategies of the parasite Giardia lamblia for evading the human immune system. Strains resistant to first-line drugs are also a challenge. Therefore, new antigiardial therapies are urgently needed. Here, we tested giardial arginine deiminase (GlADI) as a target against giardiasis. GlADI belongs to an essential pathway in Giardia for the synthesis of ATP, which is absent in humans. In silico docking with six thiol-reactive compounds was performed; four of which are approved drugs for humans. Recombinant GlADI was used in enzyme inhibition assays, and computational in silico predictions and spectroscopic studies were applied to follow the enzyme\u2019s structural disturbance and identify possible effective drugs. Inhibition by modification of cysteines was corroborated using Ellman\u2019s method. The efficacy of these drugs on parasite viability was assayed on Giardia trophozoites, along with the inhibition of the endogenous GlADI. The most potent drug against GlADI was assayed on Giardia encystment. The tested drugs inhibited the recombinant GlADI by modifying its cysteines and, potentially, by altering its 3D structure. Only rabeprazole and omeprazole decreased trophozoite survival by inhibiting endogenous GlADI, while rabeprazole also decreased the Giardia encystment rate. These findings demonstrate the potential of GlADI as a target against giardiasis.<\/jats:p>","DOI":"10.3390\/ijms22179491","type":"journal-article","created":{"date-parts":[[2021,8,31]],"date-time":"2021-08-31T21:59:45Z","timestamp":1630447185000},"page":"9491","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Multilevel Approach for the Treatment of Giardiasis by Targeting Arginine Deiminase"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7468-3420","authenticated-orcid":false,"given":"Cynthia","family":"Fern\u00e1ndez-Lainez","sequence":"first","affiliation":[{"name":"Laboratorio de Errores Innatos del Metabolismo y Tamiz, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"},{"name":"Immunoendocrinology, Division of Medical Biology, Department of Pathology and Medical Biology, University of Groningen and University Medical Centre Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands"},{"name":"Posgrado en Ciencias Biol\u00f3gicas, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Ciudad de Mexico 04510, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5654-8089","authenticated-orcid":false,"given":"Ignacio","family":"de la Mora-de la Mora","sequence":"additional","affiliation":[{"name":"Laboratorio de Biomol\u00e9culas y Salud Infantil, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3018-060X","authenticated-orcid":false,"given":"Itzhel","family":"Garc\u00eda-Torres","sequence":"additional","affiliation":[{"name":"Laboratorio de Biomol\u00e9culas y Salud Infantil, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2058-5707","authenticated-orcid":false,"given":"Sergio","family":"Enr\u00edquez-Flores","sequence":"additional","affiliation":[{"name":"Laboratorio de Biomol\u00e9culas y Salud Infantil, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luis","family":"Flores-L\u00f3pez","sequence":"additional","affiliation":[{"name":"Laboratorio de Biomol\u00e9culas y Salud Infantil, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"},{"name":"CONACYT-Instituto Nacional de Pediatr\u00eda, Secretar\u00eda de Salud, Ciudad de Mexico 04530, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9885-9635","authenticated-orcid":false,"given":"Pedro","family":"Guti\u00e9rrez-Castrell\u00f3n","sequence":"additional","affiliation":[{"name":"Hospital General Dr. Manuel Gea Gonz\u00e1lez, Ciudad de Mexico 14080, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8675-5671","authenticated-orcid":false,"given":"Lilian","family":"Y\u00e9pez-Mulia","sequence":"additional","affiliation":[{"name":"Unidad de Investigaci\u00f3n M\u00e9dica en Enfermedades Infecciosas y Parasitarias, UMAE Hospital de Pediatr\u00eda, Centro M\u00e9dico Siglo XXI, Instituto Mexicano del Seguro Social, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Felix","family":"Matadamas-Mart\u00ednez","sequence":"additional","affiliation":[{"name":"Unidad de Investigaci\u00f3n M\u00e9dica en Enfermedades Infecciosas y Parasitarias, UMAE Hospital de Pediatr\u00eda, Centro M\u00e9dico Siglo XXI, Instituto Mexicano del Seguro Social, Ciudad de Mexico 06720, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9618-2408","authenticated-orcid":false,"given":"Paul","family":"de Vos","sequence":"additional","affiliation":[{"name":"Immunoendocrinology, Division of Medical Biology, Department of Pathology and Medical Biology, University of Groningen and University Medical Centre Groningen, Hanzeplein 1, 9700 RB Groningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0470-1069","authenticated-orcid":false,"given":"Gabriel","family":"L\u00f3pez-Vel\u00e1zquez","sequence":"additional","affiliation":[{"name":"Laboratorio de Biomol\u00e9culas y Salud Infantil, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,8,31]]},"reference":[{"key":"ref_1","first-page":"586438","article-title":"Peptidylarginine deiminase inhibition abolishes the production of large extracellular vesicles from Giardia intestinalis, affecting host-pathogen interactions by hindering adhesion to host cells","volume":"10","author":"Gavinho","year":"2019","journal-title":"Biorxiv"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.biochi.2018.10.008","article-title":"Epigenetics in the early divergent eukaryotic Giardia duodenalis: An update","volume":"156","year":"2019","journal-title":"Biochimie"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0166-6851(80)90037-7","article-title":"Energy metabolism of the anaerobic protozoon Giardia lamblia","volume":"1","author":"Lindmark","year":"1980","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1016\/0020-7519(90)90133-8","article-title":"The arginine dihydrolase pathway is present in Giardia intestinalis","volume":"20","author":"Schofield","year":"1990","journal-title":"Int. J. Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/0166-6851(92)90197-R","article-title":"The pathway of arginine catabolism in Giardia intestinalis","volume":"51","author":"Schofield","year":"1992","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1128\/CMR.14.3.447-475.2001","article-title":"Biology of Giardia lamblia","volume":"14","author":"Adam","year":"2001","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2012\/980829","article-title":"Reconstruction of sugar metabolic pathways of Giardia lamblia","volume":"2012","author":"Han","year":"2012","journal-title":"Int. J. Proteom."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/0305-0491(92)90116-9","article-title":"The uptake and conversion of l-[U14C-] aspartate and l-[U14C-] alanine to 14CO2 by intact trophozoites of Giardia duodenalis","volume":"102","author":"Mendis","year":"1992","journal-title":"Comp. Biochem. Physiol. B Comp. Biochem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.bbagen.2015.10.021","article-title":"Proton pump inhibitors drastically modify triosephosphate isomerase from Giardia lamblia at functional and structural levels, providing molecular leads in the design of new antigiardiasic drugs","volume":"1860","year":"2016","journal-title":"Biochim. Et Biophys. Acta (BBA)-Gen. Subj."},{"key":"ref_10","first-page":"1","article-title":"On the molecular and cellular effects of omeprazole to further support its effectiveness as an antigiardial drug","volume":"9","author":"Ridaura","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7072","DOI":"10.1128\/AAC.02900-14","article-title":"Giardial Triosephosphate Isomerase as possible target of the cytotoxic effect of omeprazole in Giardia","volume":"58","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/bs.apar.2019.11.002","article-title":"Transcriptomic and proteomic analyses of Giardia intestinalis: Intestinal epithelial cell interactions","volume":"107","year":"2020","journal-title":"Adv. Parasitol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1921","DOI":"10.2147\/IDR.S141468","article-title":"Treatment-refractory giardiasis: Challenges and solutions","volume":"11","author":"Lalle","year":"2018","journal-title":"Infect. Drug Resist."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"7351","DOI":"10.1016\/j.bmcl.2011.10.028","article-title":"Antiprotozoal activity of proton-pump inhibitors","volume":"21","author":"Castillo","year":"2011","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_15","first-page":"1","article-title":"Synthesis and biological evaluation in vitro and in silico of N-propionyl-N\u2032-benzeneacylhydrazone derivatives as cruzain inhibitors of Trypanosoma cruzi","volume":"24","author":"Rivera","year":"2020","journal-title":"Mol. Divers."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.ijpddr.2016.07.003","article-title":"Evaluation of Giardia lamblia thioredoxin reductase as drug activating enzyme and as drug target","volume":"6","author":"Leitsch","year":"2016","journal-title":"Int. J. Parasitol. Drugs Drug Resist."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"583","DOI":"10.2174\/0929867327666200303170000","article-title":"Recent advances in the development of broad-spectrum antiprotozoal agents","volume":"28","author":"Banik","year":"2021","journal-title":"Curr. Med. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1080\/07391102.2015.1051115","article-title":"Insights into the structure and inhibition of Giardia intestinalis arginine deiminase: Homology modeling, docking, and molecular dynamics studies","volume":"34","author":"Castillo","year":"2016","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.bioorg.2009.06.001","article-title":"Mechanisms of catalysis and inhibition operative in the arginine deiminase from the human pathogen Giardia lamblia","volume":"37","author":"Li","year":"2009","journal-title":"Bioorg. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1016\/j.pt.2011.06.003","article-title":"Cyst and encystment in protozoan parasites: Optimal targets for new life-cycle interrupting strategies?","volume":"27","author":"Carrero","year":"2011","journal-title":"Trends Parasitol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2930","DOI":"10.1242\/jcs.026963","article-title":"Arginine deiminase has multiple regulatory roles in the biology of Giardia lamblia","volume":"121","author":"Touz","year":"2008","journal-title":"J. Cell Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"13330","DOI":"10.1021\/acs.jmedchem.0c00910","article-title":"Anti-Giardia Drug Discovery: Current Status and Gut Feelings","volume":"63","author":"Riches","year":"2020","journal-title":"J. Med. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.apar.2019.07.001","article-title":"Unusual proteins in Giardia duodenalis and their role in survival","volume":"106","author":"Feliziani","year":"2019","journal-title":"Adv. Parasitol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1675","DOI":"10.1016\/S1359-6446(05)03624-X","article-title":"Predicting protein druggability","volume":"10","author":"Hajduk","year":"2005","journal-title":"Drug Discov. Today"},{"key":"ref_25","first-page":"13","article-title":"Primary structure and biochemical properties of a variant-specific surface protein of Giardia","volume":"86","author":"Papanastasiou","year":"1997","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1002\/prot.20097","article-title":"An unusual triosephosphate isomerase from the early divergent eukaryote Giardia lamblia","volume":"55","author":"Cabrera","year":"2004","journal-title":"Proteins"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1016\/j.jmb.2006.10.053","article-title":"Disulfide bridges in the mesophilic triosephosphate isomerase from Giardia lamblia are related to oligomerization and activity","volume":"365","author":"Diaz","year":"2007","journal-title":"J. Mol. Biol."},{"key":"ref_28","unstructured":"Ladokhin, A.S. (2006). Fluorescence spectroscopy in peptide and protein analysis. Encycl. Anal. Chem. Appl. Theory Instrum."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1493","DOI":"10.1016\/j.bbapap.2008.05.016","article-title":"Unraveling the mechanisms of tryptophan fluorescence quenching in the triosephosphate isomerase from Giardia lamblia","volume":"1784","author":"Peon","year":"2008","journal-title":"Biochim. Et Biophys. Acta (BBA)-Proteins Proteom."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1358\/dnp.2009.22.2.1343228","article-title":"The value of drug repositioning in the current pharmaceutical market","volume":"22","author":"Tobinick","year":"2009","journal-title":"Drug News Perspect."},{"key":"ref_31","first-page":"9349063","article-title":"Role of the Synthetic B1 Vitamin Sulbutiamine on Health","volume":"2020","author":"Bettendorff","year":"2020","journal-title":"J. Nutr. Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"8591","DOI":"10.1038\/s41598-018-26845-z","article-title":"First characterization of a microsporidial triosephosphate isomerase and the biochemical mechanisms of its inactivation to propose a new druggable target","volume":"8","author":"Nava","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.ejphar.2007.10.066","article-title":"Mechanism of action of the disease-modifying anti-arthritic thiol agents D-penicillamine and sodium aurothiomalate: Restoration of cellular free thiols and sequestration of reactive aldehydes","volume":"580","author":"Wood","year":"2008","journal-title":"Eur. J. Pharmacol."},{"key":"ref_34","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":"Morrison","year":"2007","journal-title":"Science"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1093\/nar\/28.1.235","article-title":"The Protein Data Bank","volume":"28","author":"Berman","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/0263-7855(96)00018-5","article-title":"VMD: Visual molecular dynamics","volume":"14","author":"Humphrey","year":"1996","journal-title":"J. Mol. Graph."},{"key":"ref_37","first-page":"1605","article-title":"A visualization system for exploratory research and analysis. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE","volume":"25","author":"Chimera","year":"2004","journal-title":"J. Comput. Chem."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2555","DOI":"10.1007\/s00436-018-5944-1","article-title":"Giardia lamblia: Identification of molecules that contribute to direct mast cell activation","volume":"117","year":"2018","journal-title":"Parasitol. Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2411","DOI":"10.1002\/pro.5560041120","article-title":"How to measure and predict the molar absorption coefficient of a protein","volume":"4","author":"Pace","year":"1995","journal-title":"Protein Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2615","DOI":"10.1016\/S0021-9258(17)40503-5","article-title":"Arginine deiminase from Mycoplasma arthritidis. Evidence for multiple forms","volume":"252","author":"Weickmann","year":"1977","journal-title":"J. Biol. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2711","DOI":"10.1002\/prot.23100","article-title":"Determining the molecular mechanism of inactivation by chemical modification of triosephosphate isomerase from the human parasite Giardia lamblia: A study for antiparasitic drug design","volume":"79","author":"Mora","year":"2011","journal-title":"Proteins Struct. Funct. Bioinform."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/0003-9861(58)90014-6","article-title":"A colorimetric method for determining low concentrations of mercaptans","volume":"74","author":"Ellman","year":"1958","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3516","DOI":"10.1128\/iai.58.11.3516-3522.1990","article-title":"Excystation of in vitro-derived Giardia lamblia cysts","volume":"58","author":"Boucher","year":"1990","journal-title":"Infect. Immun."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/17\/9491\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:53:57Z","timestamp":1760165637000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/17\/9491"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,31]]},"references-count":43,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2021,9]]}},"alternative-id":["ijms22179491"],"URL":"https:\/\/doi.org\/10.3390\/ijms22179491","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,8,31]]}}}