{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T01:49:35Z","timestamp":1783734575427,"version":"3.55.0"},"reference-count":52,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2020,4,2]],"date-time":"2020-04-02T00:00:00Z","timestamp":1585785600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2016\/06034-2"],"award-info":[{"award-number":["2016\/06034-2"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["308351\/2013-4"],"award-info":[{"award-number":["308351\/2013-4"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["404769\/2018-7"],"award-info":[{"award-number":["404769\/2018-7"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]},{"name":"UK Research and Innovation via the Global Challenges Research Fund under grant agreement \u2018A Global Network for Neglected Tropical Diseases\u2019","award":["MR\/P027989\/1"],"award-info":[{"award-number":["MR\/P027989\/1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Trypanosoma cruzi is the aetiologic agent of Chagas disease, which affects people in the Americas and worldwide. The parasite has a complex life cycle that alternates among mammalian hosts and insect vectors. During its life cycle, T. cruzi passes through different environments and faces nutrient shortages. It has been established that amino acids, such as proline, histidine, alanine, and glutamate, are crucial to T. cruzi survival. Recently, we described that T. cruzi can biosynthesize glutamine from glutamate and\/or obtain it from the extracellular environment, and the role of glutamine in energetic metabolism and metacyclogenesis was demonstrated. In this study, we analysed the effect of glutamine analogues on the parasite life cycle. Here, we show that glutamine analogues impair cell proliferation, the developmental cycle during the infection of mammalian host cells and metacyclogenesis. Taken together, these results show that glutamine is an important metabolite for T. cruzi survival and suggest that glutamine analogues can be used as scaffolds for the development of new trypanocidal drugs. These data also reinforce the supposition that glutamine metabolism is an unexplored possible therapeutic target.<\/jats:p>","DOI":"10.3390\/molecules25071628","type":"journal-article","created":{"date-parts":[[2020,4,3]],"date-time":"2020-04-03T05:55:16Z","timestamp":1585893316000},"page":"1628","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Glutamine Analogues Impair Cell Proliferation, the Intracellular Cycle and Metacyclogenesis in Trypanosoma cruzi"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1215-8553","authenticated-orcid":false,"given":"Rodolpho Ornitz Oliveira","family":"Souza","sequence":"first","affiliation":[{"name":"Laboratory of Biochemistry of Tryps\u2013LaBTryps, Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, Av. Prof. Lineu Prestes 1374, 05508000 Sao Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcell","family":"Crispim","sequence":"additional","affiliation":[{"name":"Laboratory of Biochemistry of Tryps\u2013LaBTryps, Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, Av. Prof. Lineu Prestes 1374, 05508000 Sao Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4528-4732","authenticated-orcid":false,"given":"Ariel Mariano","family":"Silber","sequence":"additional","affiliation":[{"name":"Laboratory of Biochemistry of Tryps\u2013LaBTryps, Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, Av. Prof. Lineu Prestes 1374, 05508000 Sao Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2959-1484","authenticated-orcid":false,"given":"Fl\u00e1via Silva","family":"Damasceno","sequence":"additional","affiliation":[{"name":"Laboratory of Biochemistry of Tryps\u2013LaBTryps, Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, Av. Prof. Lineu Prestes 1374, 05508000 Sao Paulo, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,4,2]]},"reference":[{"key":"ref_1","unstructured":"WHO (2018). WHO|Chagas Disease (American Trypanosomiasis), WHO. Available online: http:\/\/www.who.int\/chagas\/en."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"170050","DOI":"10.1038\/sdata.2017.50","article-title":"The contemporary distribution of Trypanosoma cruzi infection in humans, alternative hosts and vectors","volume":"4","author":"Browne","year":"2017","journal-title":"Sci. Data"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2074","DOI":"10.2174\/138161211796904786","article-title":"The uniqueness of the Trypanosoma cruzi mitochondrion: Opportunities to identify new drug target for the treatment of Chagas disease","volume":"17","author":"Mantilla","year":"2011","journal-title":"Curr. Pharm. Des."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/S0140-6736(17)31612-4","article-title":"Chagas disease","volume":"391","author":"Molina","year":"2018","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"166","DOI":"10.3389\/fpubh.2019.00166","article-title":"Chagas disease: From discovery to a worldwide health problem","volume":"7","author":"Lidani","year":"2019","journal-title":"Front. Public Health"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/j.actatropica.2009.10.023","article-title":"Specific chemotherapy of Chagas disease: Relevance, current limitations and new approaches","volume":"115","author":"Urbina","year":"2010","journal-title":"Acta Trop."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.idc.2012.03.002","article-title":"American trypanosomiasis (Chagas disease)","volume":"26","author":"Rassi","year":"2012","journal-title":"Infect. Dis. Clin. N. Am."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1882","DOI":"10.1056\/NEJMra1410150","article-title":"Chagas\u2019 Disease","volume":"373","author":"Bern","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Marchese, L., Nascimento, J., Damasceno, F., Bringaud, F., Michels, P., and Silber, A. (2018). The uptake and metabolism of amino acids, and their unique role in the biology of pathogenic trypanosomatids. Pathogens, 7.","DOI":"10.3390\/pathogens7020036"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1007\/978-1-4899-1651-8_7","article-title":"Energy metabolism in Trypanosoma cruzi","volume":"18","author":"Cazzulo","year":"1992","journal-title":"Subcell Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"53","DOI":"10.2174\/1568005053174636","article-title":"Amino acid metabolic routes in Trypanosoma cruzi: Possible therapeutic targets against Chagas\u2019 disease","volume":"5","author":"Silber","year":"2005","journal-title":"Curr. Drug Targets Infect. Disord."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"8964","DOI":"10.1074\/jbc.M117.778522","article-title":"Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes","volume":"292","author":"Barison","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.molbiopara.2018.07.007","article-title":"L-Glutamine uptake is developmentally regulated and is involved in metacyclogenesis in Trypanosoma cruzi","volume":"224","author":"Damasceno","year":"2018","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Paes, L.S., Suarez Mantilla, B., Zimbres, F.M., Pral, E.M., Diogo de Melo, P., Tahara, E.B., Kowaltowski, A.J., Elias, M.C., and Silber, A.M. (2013). Proline dehydrogenase regulates redox state and respiratory metabolism in Trypanosoma cruzi. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069419"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3023","DOI":"10.1128\/IAI.00138-09","article-title":"Use of L-proline and ATP production by Trypanosoma cruzi metacyclic forms as requirements for host cell invasion","volume":"77","author":"Martins","year":"2009","journal-title":"Infect. Immun."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/0305-0491(76)90318-7","article-title":"Proline metabolism in Trypanosoma cruzi epimastigotes","volume":"55","author":"Sylvester","year":"1976","journal-title":"Comp. Biochem. Physiol. B"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1007\/s10863-016-9665-9","article-title":"The active transport of histidine and its role in ATP production in Trypanosoma cruzi","volume":"48","author":"Damasceno","year":"2016","journal-title":"J. Bioenerg. Biomembr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7767","DOI":"10.1074\/jbc.M114.574525","article-title":"Role of \u03941-pyrroline-5-carboxylate dehydrogenase supports mitochondrial metabolism and host-cell invasion of Trypanosoma cruzi","volume":"290","author":"Mantilla","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"566","DOI":"10.1111\/j.1550-7408.1999.tb05132.x","article-title":"L-arginine uptake and L-phosphoarginine synthesis in Trypanosoma cruzi","volume":"46","author":"Pereira","year":"1999","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/S0014-5793(02)03157-5","article-title":"Arginine metabolism in Trypanosoma cruzi is coupled to parasite stage and replication","volume":"526","author":"Pereira","year":"2002","journal-title":"FEBS Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/S0014-5793(03)01171-2","article-title":"Arginine kinase overexpression improves Trypanosoma cruzi survival capability","volume":"554","author":"Pereira","year":"2003","journal-title":"FEBS Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1016\/0166-6851(85)90073-8","article-title":"In vitro differentiation of Trypanosoma cruzi under chemically defined conditions","volume":"16","author":"Contreras","year":"1985","journal-title":"Mol. Biochem Parasitol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1111\/j.1462-5822.2004.00397.x","article-title":"L-proline is essential for the intracellular differentiation of Trypanosoma cruzi","volume":"6","author":"Tonelli","year":"2004","journal-title":"Cell Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1111\/j.1550-7408.1989.tb01063.x","article-title":"Some factors inducing formation of metacyclic stages of Trypanosoma cruzi","volume":"36","author":"Homsy","year":"1989","journal-title":"J. Protozool."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.molbiopara.2009.07.006","article-title":"Glucose uptake in the mammalian stages of Trypanosoma cruzi","volume":"168","author":"Silber","year":"2009","journal-title":"Mol. Biochem Parasitol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1016\/S0166-6851(02)00277-3","article-title":"Regulatory volume decrease in Trypanosoma cruzi involves amino acid efflux and changes in intracellular calcium","volume":"126","author":"Rohloff","year":"2003","journal-title":"Mol. Biochem Parasitol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"486928","DOI":"10.4061\/2011\/486928","article-title":"The Involvement of Glutamate Metabolism in the Resistance to Thermal, Nutritional, and Oxidative Stress in Trypanosoma cruzi","volume":"2011","author":"Magdaleno","year":"2011","journal-title":"Enzym. Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Magdaleno, A., Ahn, I.Y., Paes, L.S., and Silber, A.M. (2009). Actions of a proline analogue, L-thiazolidine-4-carboxylic acid (T4C), on Trypanosoma cruzi. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0004534"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Saye, M., Miranda, M.R., di Girolamo, F., de los Milagros Camara, M., and Pereira, C.A. (2014). Proline modulates the Trypanosoma cruzi resistance to reactive oxygen species and drugs through a novel D, L-proline transporter. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0092028"},{"key":"ref_30","first-page":"1","article-title":"Physiological role of glutaminase activity in Saccharomyces cerevisiae","volume":"133","author":"Soberon","year":"1987","journal-title":"J. Gen. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"82","DOI":"10.4161\/rna.8.1.13864","article-title":"Bacterial aptamers that selectively bind glutamine","volume":"8","author":"Ames","year":"2011","journal-title":"RNA Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1038\/onc.2009.358","article-title":"Q\u2019s next: The diverse functions of glutamine in metabolism, cell biology and cancer","volume":"29","author":"DeBerardinis","year":"2010","journal-title":"Oncogene"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"7347","DOI":"10.1074\/jbc.270.13.7347","article-title":"Biochemical characterization of human GMP synthetase","volume":"270","author":"Nakamura","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"23450","DOI":"10.1074\/jbc.270.40.23450","article-title":"The glutamine hydrolysis function of human GMP synthetase. Identification of an essential active site cysteine","volume":"270","author":"Nakamura","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"7891","DOI":"10.1021\/bi990871p","article-title":"The amidotransferase family of enzymes: Molecular machines for the production and delivery of ammonia","volume":"38","author":"Raushel","year":"1999","journal-title":"Biochemistry"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Crispim, M., Damasceno, F.S., Hernandez, A., Barison, M.J., Pretto Sauter, I., Souza Pavani, R., Santos Moura, A., Pral, E.M.F., Cortez, M., and Elias, M.C. (2018). The glutamine synthetase of Trypanosoma cruzi is required for its resistance to ammonium accumulation and evasion of the parasitophorous vacuole during host-cell infection. PLoS Negl. Trop. Dis., 12.","DOI":"10.1371\/journal.pntd.0006170"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1038\/cdd.2009.44","article-title":"Guidelines for the use and interpretation of assays for monitoring cell death in higher eukaryotes","volume":"16","author":"Galluzzi","year":"2009","journal-title":"Cell Death Differ."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1016\/j.pt.2010.06.008","article-title":"Programmed cell death in unicellular parasites: A prerequisite for sustained infection?","volume":"26","author":"Luder","year":"2010","journal-title":"Trends Parasitol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"O\u2019Rourke, R.W., Meyer, K.A., Gaston, G., White, A.E., Lumeng, C.N., and Marks, D.L. (2013). Hexosamine biosynthesis is a possible mechanism underlying hypoxia\u2019s effects on lipid metabolism in human adipocytes. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0071165"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"H815","DOI":"10.1152\/ajpheart.00756.2008","article-title":"The hexosamine biosynthesis inhibitor azaserine prevents endothelial inflammation and dysfunction under hyperglycemic condition through antioxidant effects","volume":"296","author":"Rajapakse","year":"2009","journal-title":"Am. J. Physiol. Hear. Circ. Physiol."},{"key":"ref_41","first-page":"205","article-title":"Some aspects of the purine metabolism of Trypanosoma cruzi in tissue culture","volume":"4","author":"Yoneda","year":"1971","journal-title":"Rev. Bras. Pesqui. Med. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1867","DOI":"10.1016\/0006-2952(65)90277-7","article-title":"Sensitivity of Trypanosoma equiperdum to the action of tumor-inhibitory antibiotics in vitro","volume":"14","author":"Jaffe","year":"1965","journal-title":"Biochem. Pharmacol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/0006-2952(65)90190-5","article-title":"Effect of Azaserine on the Incorporation of 14-C-Labeled Purines and pyrimidines into acid-soluble and nucleic acid fractions of Trypanosoma equiperdum","volume":"14","author":"Momparler","year":"1965","journal-title":"Biochem. Pharmacol."},{"key":"ref_44","first-page":"327","article-title":"Acivicin. An antitumor antibiotic","volume":"4","author":"Poster","year":"1981","journal-title":"Cancer Clin. Trials"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1039\/C4SC02339K","article-title":"Target discovery of acivicin in cancer cells elucidates its mechanism of growth inhibition","volume":"6","author":"Kreuzer","year":"2014","journal-title":"Chem. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1002\/cmdc.201700118","article-title":"An Improved Model of the Trypanosoma brucei CTP Synthetase Glutaminase Domain-Acivicin Complex","volume":"12","author":"Dawson","year":"2017","journal-title":"ChemMedChem"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1006","DOI":"10.1111\/mmi.13083","article-title":"GMP synthase is essential for viability and infectivity of Trypanosoma brucei despite a redundant purine salvage pathway","volume":"97","author":"Li","year":"2015","journal-title":"Mol. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Damasceno, F.S., Barison, M.J., Pral, E.M., Paes, L.S., and Silber, A.M. (2014). Memantine, an antagonist of the NMDA glutamate receptor, affects cell proliferation, differentiation and the intracellular cycle and induces apoptosis in Trypanosoma cruzi. PLoS Negl. Trop Dis., 8.","DOI":"10.1371\/journal.pntd.0002717"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1016\/j.pt.2006.08.010","article-title":"Death of a trypanosome: A selfish altruism","volume":"22","author":"Duszenko","year":"2006","journal-title":"Trends Parasitol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1038\/sj.cdd.4401564","article-title":"Prostaglandin D2 induces programmed cell death in Trypanosoma brucei bloodstream form","volume":"12","author":"Figarella","year":"2005","journal-title":"Cell Death Differ."},{"key":"ref_51","first-page":"93","article-title":"Growth and differentiation in Trypanosoma cruzi. I. Origin of metacyclic trypanosomes in liquid media","volume":"6","author":"Camargo","year":"1964","journal-title":"Rev. Inst. Med. Trop. Sao Paulo"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0022-1759(83)90303-4","article-title":"Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays","volume":"65","author":"Mosmann","year":"1983","journal-title":"J. Immunol. Methods"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/7\/1628\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:14:32Z","timestamp":1760174072000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/25\/7\/1628"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,4,2]]},"references-count":52,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["molecules25071628"],"URL":"https:\/\/doi.org\/10.3390\/molecules25071628","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,4,2]]}}}