Skip to main content Accessibility help

The effect of iron on Trichomonas vaginalis TvCP2: a cysteine proteinase found in vaginal secretions of trichomoniasis patients

  • Luis Alberto Rivera-Rivas (a1), Sebastián Lorenzo-Benito (a1), Diana Belén Sánchez-Rodríguez (a1), Jesús FT Miranda-Ozuna (a1), Esly Alejandra Euceda-Padilla (a1), Jaime Ortega-López (a2), Bibiana Chávez-Munguía (a1), Anel Lagunes-Guillén (a1), Beatriz Velázquez-Valassi (a3), Lidia Jasso-Villazul (a4) and Rossana Arroyo (a1)...


Trichomonas vaginalis (Tv) induces host cell damage through cysteine proteinases (CPs) modulated by iron. An immunoproteomic analysis showed that trichomoniasis patient sera recognize various CPs, also some of them are present in vaginal washes (VWs). Thus, the goal of this work was to determine whether TvCP2 is expressed during infection and to assess the effect of iron on TvCP2 expression, localization and contribution to in vitro cellular damage. Western-blotting (WB) assays using TvCP2r and vaginitis patient serum samples showed that 6/9 Tv (+) but none of the Tv (−) patient sera recognized TvCP2r. WB using an anti-TvCP2r antibody and VWs from the same patients showed that in all of the Tv (+) but none of the Tv (−) VWs, the anti-TvCP2r antibody detected a 27 kDa protein band that corresponded to the mature TvCP2, which was confirmed by mass spectrometry analysis. Iron decreased the amount of TvCP2 mRNA and the protein localized on the parasite surface and cytoplasmic vesicles concomitant with the cytotoxic effect of TvCP2 on HeLa cells. Parasites pretreated with the anti-TvCP2r antibody also showed reduced levels of cytotoxicity and apoptosis induction in HeLa cell monolayers. In conclusion, these results show that TvCP2 is expressed during trichomonal infection and plays an important role in the in vitro HeLa cell cytotoxic damage under iron-restricted conditions.


Corresponding author

Author for correspondence: Rossana Arroyo, E-mail:


Hide All
Adler, J and Parmryd, I (2010) Quantifying colocalization by correlation: the Pearson correlation coefficient is superior to the Mander's overlap coefficient. Cytometry Part A, 77, 732742. doi: 10.1002/cyto.a.20896.
Alderete, JF, Provenzano, D and Lehker, MW (1995) Iron mediates Trichomonas vaginalis resistance to complement lysis. Microbial Pathogens 19, 93103.10.1006/mpat.1995.0049
Alvarez-Sánchez, ME, Ávila-González, L, Becerril-García, C, Fattel-Facenda, LV, Ortega-López, J and Arroyo, R (2000) A novel cysteine proteinase (CP65) of Trichomonas vaginalis involved in cytotoxicity. Microbial Pathogens 28, 193202.10.1006/mpat.1999.0336
Alvarez-Sánchez, ME, Solano-González, E, Yáñez-Gómez, C and Arroyo, R (2007) Negative iron regulation of the CP65 cysteine proteinase cytotoxicity in Trichomonas vaginalis. Microbes and Infection 9, 15971605.10.1016/j.micinf.2007.09.011
Arroyo, R and Alderete, JF (1989) Trichomonas vaginalis surface proteinase activity is necessary for adherence to epithelial cells. Infection and Immunity 57, 29912997.10.1128/IAI.57.10.2991-2997.1989
Arroyo, R and Alderete, JF (1995) Two Trichomonas vaginalis surface proteinases bind to host epithelial cells and are related to levels of cytoadherence and cytotoxicity. Archives of Medical Research 26, 279285.
Arroyo, R, Cárdenas-Guerra, RE, Figueroa-Angulo, EE, Puente-Rivera, J, Zamudio-Prieto, O and Ortega-López, J (2015) Trichomonas vaginalis cysteine proteinases: iron response in gene expression and proteolytic activity. Biomed Research International 2015, 946787. doi: 10.1155/2015/946787.
Benchimol, M (2004) Trichomonads under microscopy. Microscopy and Microanalysis 10, 528550.10.1017/S1431927604040905
Cárdenas-Guerra, RE, Arroyo, R, Rosa de Andrade, I, Benchimol, M and Ortega-López, J (2013) The iron-induced cysteine proteinase TvCP4 plays a key role in Trichomonas vaginalis haemolysis. Microbes and Infection 15, 958968.10.1016/j.micinf.2013.09.002
Cárdenas-Guerra, RE, Ortega-López, J, Flores-Pucheta, CI, Benítez-Cardoza, CG and Arroyo, R (2015) The recombinant prepro region of TvCP4 is an inhibitor of cathepsin L-like cysteine proteinases of Trichomonas vaginalis that inhibits trichomonal haemolysis. International Journal of Biochemistry and Cell Biology 59, 7383.10.1016/j.biocel.2014.12.001
Carlton, JM, Hirt, RP, Silva, JC, Delcher, AL, Schatz, M, Zhao, Q, Wortman, JR, Bidwell, SL, Alsmark, UC, Besteiro, S, Sicheritz-Ponten, T, Noel, CJ, Dacks, JB, Foster, PG, Simillion, C, Van de Peer, Y, Miranda-Saavedra, D, Barton, GJ, Westrop, GD, Müller, S, Dessi, D, Fiori, PL, Ren, Q, Paulsen, I, Zhang, H, Bastida-Corcuera, FD, Simoes-Barbosa, A, Brown, MT, Hayes, RD, Mukherjee, M, Okumura, CY, Schneider, R, Smith, AJ, Vanacova, S, Villalvazo, M, Haas, BJ, Pertea, M, Feldblyum, TV, Utterback, TR, Shu, CL, Osoegawa, K, de Jong, PJ, Hrdy, I, Horvathova, L, Zubacova, Z, Dolezal, P, Malik, SB, Logsdon, JM Jr, Henze, K, Gupta, A, Wang, CC, Dunne, RL, Upcroft, JA, Upcroft, P, White, O, Salzberg, SL, Tang, P, Chiu, CH, Lee, YS, Embley, TM, Coombs, GH, Mottram, JC, Tachezy, J, Fraser-Liggett, CM and Johnson, PJ (2007) Draft genome sequence of the sexually transmitted pathogen Trichomonas vaginalis. Science (New York, N.Y.) 315, 207212.10.1126/science.1132894
Carvajal-Gámez, BI, Quintas-Granados, LI, Arroyo, R, Vázquez-Carrillo, LI, Ramón-Luing, LL, Carrillo-Tapia, E and Álvarez-Sánchez, ME (2014) Putrescine-dependent re-localization of TvCP39, a cysteine proteinase involved in Trichomonas vaginalis cytotoxicity. PLoS ONE 9, e107293.10.1371/journal.pone.0107293
Conrad, MD, Gorman, AW, Schillinger, JA, Fiori, PL, Arroyo, R, Malla, N, Dubey, ML, González, J, Blank, S, Secor, WE and Carlton, JM (2012) Extensive genetic diversity, unique population structure and evidence of genetic exchange in the sexually transmitted parasite Trichomonas vaginalis. PLoS Neglected Tropical Diseases 6, e1573.10.1371/journal.pntd.0001573
Dailey, DC, Chang, TH and Alderete, JF (1990) Characterization of Trichomonas vaginalis haemolysis. Parasitology 101, 171175.10.1017/S0031182000063204
De Jesus, JB, Ferreira, MA, Cuervo, P, Britto, C, Silva-Filho, FC and Meyer-Fernandes, JR (2006) Iron modulates ecto-phosphohydrolase activities in pathogenic trichomonads. Parasitoloy Internacional 55, 285290.
De Jesus, JB, Cuervo, P, Junqueira, M, Britto, C, Silva-Filho, FC, Soares, MJ, Cupolillo, E, Fernandes, O and Domont, GB (2007) A further proteomic study on the effect of iron in the human pathogen Trichomonas vaginalis. Proteomics 7, 19611972.10.1002/pmic.200600797
Diamond, LS (1957) The establishment of various trichomonads of animals and man in axenic cultures. Journal of Parasitology 43, 488490.10.2307/3274682
Dias-Lopes, G, Saboia-Vahia, L, Margotti, ET, Fernandes, NS, Castro, CLF, Oliveira, FO, Peixoto, JF, Britto, C, Silva-Filho, FCE, Cuervo, P and Jesus, JB (2017) Morphologic study of the effect of iron on pseudocysts formation un Trichomonas vaginalis and its interaction with human epithelial cells. Memórias do Instituto Oswaldo Cruz 112, 664673.10.1590/0074-02760170032
Figueroa-Angulo, EE, Rendón-Gandarilla, FJ, Puente-Rivera, J, Calla-Choque, JS, Cárdenas-Guerra, RE, Ortega-López, J, Quintas-Granados, LI, Álvarez-Sánchez, ME and Arroyo, R (2012) The effects of environmental factors on the virulence of Trichomonas vaginalis. Microbes and Infection 14, 14111427.10.1016/j.micinf.2012.09.004
Fiori, PL, Rappelli, P, Rocchigiani, AM and Cappuccinelli, P (1993) Trichomonas vaginalis haemolysis: evidence of functional pores formation on red cell membranes. FEMS Microbiology 109, 1318.10.1111/j.1574-6968.1993.tb06136.x
Fiori, PL, Rappelli, P, Addis, MF, Sechi, A and Capuccinelli, P (1996) Trichomonas vaginalis haemolysis: pH regulates a contact-independent mechanism based on pore-forming proteins. Microbial Pathogens 20, 109118.10.1006/mpat.1996.0010
Fiori, PL, Rappelli, P and Addis, MF (1999) The flagellated parasite Trichomonas vaginalis: new insights into cytopathogenicity mechanisms. Microbes and Infection 1, 149156.10.1016/S1286-4579(99)80006-9
Gorrell, TE (1985) Effect of culture medium iron content on the biochemical composition and metabolism of Trichomonas vaginalis. Journal of Bacteriology 161, 12281230.10.1128/JB.161.3.1228-1230.1985
Harlow, E and Lane, D (1988) Antibodies: A Laboratory Manual. New York: Cold Spring Harbor Laboratory, pp. 298300. ISBN 0-87969-314-2.
Hernández-Gutiérrez, R, Ortega-López, J and Arroyo, R (2003) A 39-kDa cysteine proteinase CP39 from Trichomonas vaginalis, which is negatively affected by iron may be involved in trichomonal cytotoxicity. Journal of Eukaryotic Microbiology 50(Suppl), 696698.
Hernández-Gutiérrez, R, Avila González, L, Ortega-López, J, Cruz-Talonia, F, Gómez-Gutiérrez, G and Arroyo, R (2004) Trichomonas vaginalis: characterization of a 39-kDa cysteine proteinase found in patient vaginal secretions. Experimental Parasitology 107, 125135.
Hernández, HM, Marcet, R and Sarracent, J (2014) Biological roles of cysteine proteinases in the pathogenesis of Trichomonas vaginalis. Parasite 21, 54.
Horváthová, L, Šafaríková, L, Basler, M, Hrdy, I, Campo, NB, Shin, JW, Huang, KY, Huang, PJ, Lin, R, Tang, P and Tachezy, J (2012) Transcriptomic identification of iron-regulated and iron-independent gene copies within the heavily duplicated Trichomonas vaginalis genome. Genome Biological Evolution 4, 10171029.
Kummer, S, Hayes, GR, Gilbert, RO, Beach, DH, Lucas, JJ and Singh, BN (2008) Induction of human host cell apoptosis by Trichomonas vaginalis cysteine proteases is modulated by parasite exposure to iron. Microbial Pathogens 44, 197203.
Lehker, MW and Alderete, JF (1992) Iron regulates growth of Trichomonas vaginalis and the expression of immunogenic trichomonad proteins. Molecular Microbiology 6, 123132.
Lehker, MW and Alderete, JF (2000) Biology of trichomonosis. Current Opinion in Infectious Diseases 13, 745.
León-Sicairos, CR, León-Félix, J and Arroyo, R (2004) Tvcp12: a novel Trichomonas vaginalis cathepsin L-like cysteine proteinase encoding-gene. Microbiology (Reading, England) 150, 11311138.
Lubick, KJ and Burgess, DE (2004) Purification and analysis of a phospholipase A2-like lytic factor of Trichomonas vaginalis. Infection and Immunity 72, 12841290.
Madico, G, Quinn, TC, Rompalo, A, McKee, KT and Gaydós, CA (1998) Diagnosis of Trichomonas vaginalis infection by using vaginal swab samples. Journal of Clinical Microbiology 36, 32053210.
Mallinson, DJ, Lockwood, BC, Coombs, GH and North, MJ (1994) Identification and molecular cloning of four cysteine proteinase genes from the pathogenic protozoon Trichomonas vaginalis. Microbiology (Reading, England) 140, 27252735.10.1099/00221287-140-10-2725
Mendoza-López, MR, Becerril-García, C, Fattel-Facenda, LV, Ávila-González, L, et al. (2000) CP30, A cysteine proteinase involved in Trichomonas vaginalis cytoadherence. Infection and Immunity 68, 49074912.
Meza-Cervantez, P, González-Robles, A, Cárdenas-Guerra, RE, Ortega-López, J, Saavedra, E, Pineda, E and Arroyo, R (2011) Pyruvate:ferredoxin oxidoreductase (PFO) is a surface-associated cell-binding protein in Trichomonas vaginalis and is involved in trichomonal adherence to host cells. Microbiology (Reading, England) 157, 34693482.
Miranda-Ozuna, JFT, Hernández-García, MS, Brieba, LG, Benítez-Cardoza, CG, Ortega-López, J, González-Robles, A and Arroyo, R (2016) The glycolytic enzyme triosephosphate isomerase of Trichomonas vaginalis is a surface-associated protein induced by glucose that functions as a laminin- and fibronectin-binding protein. Infection and Immunity 84, 28782894.
Miranda-Ozuna, JFT, Rivera-Rivas, LA, Cárdenas-Guerra, RE, Hernández-García, MS, Rodríguez-Cruz, S, González-Robles, A, Chávez-Munguía, B and Arroyo, R (2019) Glucose-restriction increases Trichomonas vaginalis cellular damage towards HeLa cells and proteolytic activity of cysteine proteinases (CPs), such as TvCP2. Parasitology 146, 11561166.
Neale, KA and Alderete, JF (1990) Analysis of the proteinases of representative Trichomonas vaginalis isolates. Infection and Immunity 58, 157162.
Ortíz-Estrada, G, Calderón-Salinas, V, Shibayama-Salas, M, León-Sicairos, N and De la Garza, M (2015) Binding and endocytosis of bovine hololactoferrin by the parasite Entamoeba histolytica. Biomedical Research International 2015, 375836.
Pastorek, L, Sobol, M and Hozák, P (2016) Colocalization coefficients evaluating the distribution of molecular targets in microscopy methods based on pointed patterns. Histochemical Cell Biology 146, 391406.
Petrin, D, Delgaty, K, Bhatt, R and Garber, G (1998) Clinical and microbiological aspects of Trichomonas vaginalis. Clinical Microbiology Reviews 11, 300317.
Puente-Rivera, J, Ramón-Luing, LA, Figueroa-Angulo, EE, Ortega-López, J and Arroyo, R (2014) Trichocystatin-2 (TC-2): an endogenous inhibitor of cysteine proteinases in Trichomonas vaginalis is associated with TvCP39. International Journal of Biochemistry and Cell Biology 54C, 255265.
Puente-Rivera, J, Villalpando, JL, Villalobos-Osnaya, A, Vázquez-Carrillo, LI, León-Avila, G, Ponce-Regalado, MD, López-Camarillo, C, Elizalde-Contreras, JM, Ruiz-May, E, Arroyo, R and Alvarez-Sánchez, ME (2017) The 50 kDa metalloproteinase TvMP50 is a zinc-mediated Trichomonas vaginalis virulence factor. Molecular Biochemical Parasitology 217, 3241.
Ramón-Luing, LA, Rendón-Gandarilla, FJ, Puente-Rivera, J, Ávila-González, L and Arroyo, R (2011) Identification and characterization of the immunogenic cytotoxic TvCP39 proteinase gene of Trichomonas vaginalis. International Journal of Biochemistry and Cell Biology 43, 15001511.10.1016/j.biocel.2011.07.001
Ramón-Luing, LA, Rendón-Gandarilla, FJ, Cárdenas-Guerra, RE, Rodríguez-Cabrera, N, Ortega-López, J, et al. (2010) Immunoproteomics of the active degradome to identify biomarkers for Trichomonas vaginalis. Proteomics 10, 435444.
Rendón-Gandarilla, FJ, Ramón-Luing, LA, Ortega-López, J, Rosa de Andrade, I, Benchimol, M and Arroyo, R (2013) The TvLEGU-1, a legumain-like cysteine proteinase, plays a key role in Trichomonas vaginalis cytoadherence. Biomed Research International 2013, 561979. doi: 10.1155/2013/561979.
Ryu, JS, Choi, HK, Min, DY, Ha, SE and Ahn, MH (2001) Effect of iron on the virulence of Trichomonas vaginalis. Journal of Parasitology 87, 457460.
Sánchez-Rodríguez, DB, Ortega-López, J, Cárdenas-Guerra, RE, Reséndiz-Cardiel, G, Chávez-Munguía, B, Lagunes-Guillén, A and Arroyo, R (2018) Characterization of a novel endogenous cysteine proteinase inhibitor trichocystatin-3 (TC-3), localized on the surface of Trichomonas vaginalis. International Journal of Biochemistry and Cell Biology 102, 87100.
Singh, M, Sharma, H and Singh, N (2007) Hydrogen peroxide induces apoptosis in HeLa cells through mitochondrial pathway. Mitochondrion 7, 367373.
Solano-González, E, Burrola-Barraza, E, León-Sicairos, C, Ávila-González, L, Gutiérrez-Escolano, L, Ortega-López, J and Arroyo, R (2007) The trichomonad cysteine proteinase TvCP4 transcript contains an iron-responsive element. FEBS Letters 581, 29192928.
Sommer, U, Costello, CE, Hayes, GR, Beach, DH, Gilbert, DO, et al. (2005) Identification of Trichomonas vaginalis cysteine proteases that induce apoptosis in human vaginal epithelial cells. Journal of Biological Chemistry 280, 28532860.10.1074/jbc.M501752200
Torres-Romero, JC and Arroyo, R (2009) Responsiveness of Trichomonas vaginalis to iron concentrations: evidence for a post-transcriptional iron regulation by an IRE/IRP-like system. Infection Genetics and Evolution 9, 10651074.10.1016/j.meegid.2009.06.003
Tsai, CD, Liu, HS and Tai, JH (2002) Characterization of an iron-responsive promoter in the protozoan pathogen Trichomonas vaginalis. Journal of Biological Chemistry 277, 51535162.
World Health Organization (2012) A baseline report on global sexually transmitted infection surveillance.


Type Description Title
Supplementary materials

Rivera-Rivas et al. supplementary material
Rivera-Rivas et al. supplementary material

 Unknown (1.2 MB)
1.2 MB

The effect of iron on Trichomonas vaginalis TvCP2: a cysteine proteinase found in vaginal secretions of trichomoniasis patients

  • Luis Alberto Rivera-Rivas (a1), Sebastián Lorenzo-Benito (a1), Diana Belén Sánchez-Rodríguez (a1), Jesús FT Miranda-Ozuna (a1), Esly Alejandra Euceda-Padilla (a1), Jaime Ortega-López (a2), Bibiana Chávez-Munguía (a1), Anel Lagunes-Guillén (a1), Beatriz Velázquez-Valassi (a3), Lidia Jasso-Villazul (a4) and Rossana Arroyo (a1)...


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.