Polyamines pp 309-326 | Cite as

Genetic and Biochemical Analysis of Protozoal Polyamine Transporters

  • Marie-Pierre Hasne
  • Buddy Ullman
Part of the Methods in Molecular Biology book series (MIMB, volume 720)


Polyamines are aliphatic polycations that function in key cellular processes such as growth, differentiation, and macromolecular biosynthesis. Intracellular polyamines pools are maintained from de novo synthesis and from transport of polyamines from the extracellular milieu. This acquisition of exogenous polyamines is mediated by cell surface transporter proteins. Protozoan parasites are the etiologic agents of a plethora of devastating and often fatal diseases in humans and their domestic animals. These pathogens accommodate de novo and/or salvage mechanisms for polyamine acquisition. Because of its therapeutic relevance, the polyamine biosynthetic pathway has been thoroughly investigated in many genera of protozoan parasites, but the polyamine permeation pathways have generally been ignored. Our group has now identified at the molecular level polyamine transporters from two species of protozoan parasites, Leishmania major and Trypanosoma cruzi, characterized these polytopic proteins with respect to ligand specificities and affinities, and determined the subcellular environments in which these transporters reside.

Key words

Polyamines Diamines Transporters Parasites Leishmania Trypanosoma cruzi Localization Confocal microscopy 



This work was supported in part by Grant-in-Aid #0950095G provided by the American Heart Association and by Grant AI41622 from the National Institute of Allergy and Infectious Disease.


  1. 1.
    Pegg AE, McGovern KA, Wiest L (1987) Decarboxylation of alpha-difluoromethylornithine by ornithine decarboxylase. Biochem J 241:305–307PubMedGoogle Scholar
  2. 2.
    Bacchi CJ, McCann PP (1987) Parasitic protozoa and polyamines. In: McCann PP, Pegg AE, Sjoerdsma A (eds) Inhibition of polyamine metabolism: biological significance and basis for new therapies. Academic, Orlando, FL, pp 317–344Google Scholar
  3. 3.
    Burri C, Brun R (2003) Eflornithine for the treatment of human African trypanosomiasis. Parasitol Res 90(Supp 1):S49–S52PubMedGoogle Scholar
  4. 4.
    Pepin J, Milord F (1994) The treatment of human African trypanosomiasis. Adv Parasitol 33:1–47PubMedCrossRefGoogle Scholar
  5. 5.
    Van Nieuwenhove S, Schechter PJ, Declercq J, Bone G, Burke J, Sjoerdsma A (1985) Treatment of gambiense sleeping sickness in the Sudan with oral DFMO (DL-alpha-difluoromethylornithine), an inhibitor of ornithine decarboxylase; first field trial. Trans R Soc Trop Med Hyg 79:692–698PubMedCrossRefGoogle Scholar
  6. 6.
    Bacchi CJ, Nathan HC, Yarlett N, Goldberg B, McCann PP, Bitonti AJ, Sjoerdsma A (1992) Cure of murine Trypanosoma brucei rhodesiense infections with an S-adenosylmethionine decarboxylase inhibitor. Antimicrob Agents Chemother 36:2736–2740PubMedGoogle Scholar
  7. 7.
    Bitonti AJ, Byers TL, Bush TL, Casara PJ, Bacchi CJ, Clarkson AB Jr, McCann PP, Sjoerdsma A (1990) Cure of Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense infections in mice with an irreversible inhibitor of S-adenosylmethionine decarboxylase. Antimicrob Agents Chemother 34:1485–1490PubMedGoogle Scholar
  8. 8.
    Bitonti AJ, Dumont JA, McCann PP (1986) Characterization of Trypanosoma brucei brucei S-adenosyl-L-methionine decarboxylase and its inhibition by Berenil, pentamidine and methylglyoxal bis(guanylhydrazone). Biochem J 237:685–689PubMedGoogle Scholar
  9. 9.
    Danzin C, Marchal P, Casara P (1990) Irreversi-ble inhibition of rat S-adenosylmethionine decarboxylase by 5’-([(Z)-4-amino-2-butenyl]methylamino)-5’-deoxyadenosine. Biochem Pharmacol 40:1499–1503PubMedCrossRefGoogle Scholar
  10. 10.
    Balana-Fouce R, Escribano MI, Alunda JM (1991) Leishmania infantum: polyamine biosynthesis and levels during the growth of promastigotes. Int J Biochem 23:1213–1217PubMedCrossRefGoogle Scholar
  11. 11.
    Ariyanayagam MR, Fairlamb AH (1997) Diamine auxotrophy may be a universal feature of Trypanosoma cruzi epimastigotes. Mol Biochem Parasitol 84:111–121PubMedCrossRefGoogle Scholar
  12. 12.
    Carrillo C, Cejas S, Gonzalez NS, Algranati ID (1999) Trypanosoma cruzi epimastigotes lack ornithine decarboxylase but can express a foreign gene encoding this enzyme. FEBS Lett 454:192–196PubMedCrossRefGoogle Scholar
  13. 13.
    Magagnin S, Bertran J, Werner A, Markovich D, Biber J, Palacin M, Murer H (1992) Poly(A)+ RNA from rabbit intestinal mucosa induces b0,+ and y+ amino acid transport activities in Xenopus laevis oocytes. J Biol Chem 267:15384–15390PubMedGoogle Scholar
  14. 14.
    Sanchez MA, Tryon R, Green J, Boor I, Landfear SM (2002) Six related nucleoside/nucleobase transporters from Trypanosoma brucei exhibit distinct biochemical functions. J Biol Chem 277:21499–21504PubMedCrossRefGoogle Scholar
  15. 15.
    Hasne MP, Ullman B (2005) Identification and characterization of a polyamine permease from the protozoan parasite Leishmania major. J Biol Chem 280:15188–15194PubMedCrossRefGoogle Scholar
  16. 16.
    Nishimura H, Pallardo FV, Seidner GA, Vannucci S, Simpson IA, Birnbaum MJ (1993) Kinetics of GLUT1 and GLUT4 glucose transporters expressed in Xenopus oocytes.J Biol Chem 268:8514–8520PubMedGoogle Scholar
  17. 17.
    Ha DS, Schwarz JK, Turco SJ, Beverley SM (1996) Use of the green fluorescent protein as a marker in transfected Leishmania. Mol Biochem Parasitol 77:57–64PubMedCrossRefGoogle Scholar
  18. 18.
    Wirtz E, Hartmann C, Clayton C (1994) Gene expression mediated by bacteriophage T3 and T7 RNA polymerases in transgenic trypanosomes. Nucleic Acids Res 22:3887–3894PubMedCrossRefGoogle Scholar
  19. 19.
    Kelly JM, Ward HM, Miles MA, Kendall G (1992) A shuttle vector which facilitates the expression of transfected genes in Trypanosoma cruzi and Leishmania. Nucleic Acids Res 20:3963–3969PubMedCrossRefGoogle Scholar
  20. 20.
    Montalvetti A, Rohloff P, Docampo R (2004) A functional aquaporin co-localizes with the vacuolar proton pyrophosphatase to acidocalcisomes and the contractile vacuole complex of Trypanosoma cruzi. J Biol Chem 279:38673–38682PubMedCrossRefGoogle Scholar
  21. 21.
    Arriza JL, Kavanaugh MP, Fairman WA, Wu YN, Murdoch GH, North RA, Amara SG (1993) Cloning and expression of a human neutral amino acid transporter with structural similarity to the glutamate transporter gene family. J Biol Chem 268:15329–15332PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Marie-Pierre Hasne
    • 1
  • Buddy Ullman
    • 1
  1. 1.Department of Biochemistry and Molecular BiologyOregon Health and Science UniversityPortlandUSA

Personalised recommendations