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Biochem J. Apr 15, 1999; 339(Pt 2): 363–370.
PMCID: PMC1220166

Inhibition of the peroxidative degradation of haem as the basis of action of chloroquine and other quinoline antimalarials.

Abstract

The malaria parasite feeds by degrading haemoglobin in an acidic food vacuole, producing free haem moieties as a by-product. The haem in oxyhaemoglobin is oxidized from the Fe(II) state to the Fe(III) state with the consequent production of an equimolar concentration of H2O2. We have analysed the fate of haem molecules in Plasmodium falciparum-infected erythrocytes and have found that only about one third of the haem is polymerized to form haemozoin. The remainder appears to be degraded by a non-enzymic process which leads to an accumulation of iron in the parasite. A possible route for degradation of the haem is by reacting with H2O2, and we show that, under conditions designed to resemble those found in the food vacuole, i.e., at pH5.2 in the presence of protein, free haem undergoes rapid peroxidative decomposition. Chloroquine and quinacrine are shown to be efficient inhibitors of the peroxidative destruction of haem, while epiquinine, a quinoline compound with very low antimalarial activity, has little inhibitory effect. We also show that chloroquine enhances the association of haem with membranes, while epiquinine inhibits this association, and that treatment of parasitized erythrocytes with chloroquine leads to a build-up of membrane-associated haem in the parasite. We suggest that chloroquine exerts its antimalarial activity by causing a build-up of toxic membrane-associated haem molecules that eventually destroy the integrity of the malaria parasite. We have further shown that resistance-modulating compounds, such as chlorpromazine, interact with haem and efficiently inhibit its degradation. This may explain the weak antimalarial activities of these compounds.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Foley M, Tilley L. Quinoline antimalarials: mechanisms of action and resistance and prospects for new agents. Pharmacol Ther. 1998 Jul;79(1):55–87. [PubMed]
  • Yayon A, Timberg R, Friedman S, Ginsburg H. Effects of chloroquine on the feeding mechanism of the intraerythrocytic human malarial parasite Plasmodium falciparum. J Protozool. 1984 Aug;31(3):367–372. [PubMed]
  • Eckman JR, Modler S, Eaton JW, Berger E, Engel RR. Host heme catabolism in drug-sensitive and drug-resistant malaria. J Lab Clin Med. 1977 Oct;90(4):767–770. [PubMed]
  • Slater AF, Cerami A. Inhibition by chloroquine of a novel haem polymerase enzyme activity in malaria trophozoites. Nature. 1992 Jan 9;355(6356):167–169. [PubMed]
  • Dorn A, Stoffel R, Matile H, Bubendorf A, Ridley RG. Malarial haemozoin/beta-haematin supports haem polymerization in the absence of protein. Nature. 1995 Mar 16;374(6519):269–271. [PubMed]
  • Raynes K, Foley M, Tilley L, Deady LW. Novel bisquinoline antimalarials. Synthesis, antimalarial activity, and inhibition of haem polymerisation. Biochem Pharmacol. 1996 Aug 23;52(4):551–559. [PubMed]
  • Wallace WJ, Houtchens RA, Maxwell JC, Caughey WS. Mechanism of autooxidation for hemoglobins and myoglobins. Promotion of superoxide production by protons and anions. J Biol Chem. 1982 May 10;257(9):4966–4977. [PubMed]
  • Carrell RW, Winterbourn CC, Rachmilewitz EA. Activated oxygen and haemolysis. Br J Haematol. 1975 Jul;30(3):259–264. [PubMed]
  • Fairfield AS, Abosch A, Ranz A, Eaton JW, Meshnick SR. Oxidant defense enzymes of Plasmodium falciparum. Mol Biochem Parasitol. 1988 Jul;30(1):77–82. [PubMed]
  • Gamain B, Langsley G, Fourmaux MN, Touzel JP, Camus D, Dive D, Slomianny C. Molecular characterization of the glutathione peroxidase gene of the human malaria parasite Plasmodium falciparum. Mol Biochem Parasitol. 1996 Jun;78(1-2):237–248. [PubMed]
  • Hunt NH, Stocker R. Oxidative stress and the redox status of malaria-infected erythrocytes. Blood Cells. 1990;16(2-3):499–530. [PubMed]
  • Brown SB, Dean TC, Jones P. Catalatic activity of iron(3)-centred catalysts. Role of dimerization in the catalytic action of ferrihaems. Biochem J. 1970 May;117(4):741–744. [PMC free article] [PubMed]
  • Hatzikonstantinou H, Brown SB. Catalase model systems. Decomposition of hydrogen peroxide catalysed by mesoferrihaem, deuteroferrihaem, coproferrihaem and haematoferrihaem. Biochem J. 1978 Sep 15;174(3):893–900. [PMC free article] [PubMed]
  • Raynes K, Galatis D, Cowman AF, Tilley L, Deady LW. Synthesis and activity of some antimalarial bisquinolines. J Med Chem. 1995 Jan 6;38(1):204–206. [PubMed]
  • La Greca N, Hibbs AR, Riffkin C, Foley M, Tilley L. Identification of an endoplasmic reticulum-resident calcium-binding protein with multiple EF-hand motifs in asexual stages of Plasmodium falciparum. Mol Biochem Parasitol. 1997 Nov;89(2):283–293. [PubMed]
  • Riggs A. Preparation of blood hemoglobins of vertebrates. Methods Enzymol. 1981;76:5–29. [PubMed]
  • Aley SB, Sherwood JA, Marsh K, Eidelman O, Howard RJ. Identification of isolate-specific proteins on sorbitol-enriched Plasmodium falciparum infected erythrocytes from Gambian patients. Parasitology. 1986 Jun;92(Pt 3):511–525. [PubMed]
  • Asakura T, Minakata K, Adachi K, Russell MO, Schwartz E. Denatured hemoglobin in sickle erythrocytes. J Clin Invest. 1977 Apr;59(4):633–640. [PMC free article] [PubMed]
  • Carter P. Spectrophotometric determination of serum iron at the submicrogram level with a new reagent (ferrozine). Anal Biochem. 1971 Apr;40(2):450–458. [PubMed]
  • Chou AC, Fitch CD. Control of heme polymerase by chloroquine and other quinoline derivatives. Biochem Biophys Res Commun. 1993 Aug 31;195(1):422–427. [PubMed]
  • Slater AF, Swiggard WJ, Orton BR, Flitter WD, Goldberg DE, Cerami A, Henderson GB. An iron-carboxylate bond links the heme units of malaria pigment. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):325–329. [PMC free article] [PubMed]
  • Wood PA, Eaton JW. Hemoglobin catabolism and host-parasite heme balance in chloroquine-sensitive and chloroquine-resistant Plasmodium berghei infections. Am J Trop Med Hyg. 1993 Apr;48(4):465–472. [PubMed]
  • Brown SB, Hatzikonstantinou H, Herries DG. The role of peroxide in haem degradation. A study of the oxidation of ferrihaems by hydrogen peroxide. Biochem J. 1978 Sep 15;174(3):901–907. [PMC free article] [PubMed]
  • Sullivan DJ, Jr, Gluzman IY, Goldberg DE. Plasmodium hemozoin formation mediated by histidine-rich proteins. Science. 1996 Jan 12;271(5246):219–222. [PubMed]
  • Kuzelová K, Mrhalová M, Hrkal Z. Kinetics of heme interaction with heme-binding proteins: the effect of heme aggregation state. Biochim Biophys Acta. 1997 Oct 20;1336(3):497–501. [PubMed]
  • Rosenthal PJ, Meshnick SR. Hemoglobin catabolism and iron utilization by malaria parasites. Mol Biochem Parasitol. 1996 Dec 20;83(2):131–139. [PubMed]
  • Cannon JB, Kuo FS, Pasternack RF, Wong NM, Muller-Eberhard U. Kinetics of the interaction of hemin liposomes with heme binding proteins. Biochemistry. 1984 Jul 31;23(16):3715–3721. [PubMed]
  • Yayon A, Cabantchik ZI, Ginsburg H. Identification of the acidic compartment of Plasmodium falciparum-infected human erythrocytes as the target of the antimalarial drug chloroquine. EMBO J. 1984 Nov;3(11):2695–2700. [PMC free article] [PubMed]
  • Zhang Y, Hempelmann E. Lysis of malarial parasites and erythrocytes by ferriprotoporphyrin IX-chloroquine and the inhibition of this effect by proteins. Biochem Pharmacol. 1987 Apr 15;36(8):1267–1273. [PubMed]
  • Blauer G, Akkawi M. Investigations of B- and beta-hematin. J Inorg Biochem. 1997 May 1;66(2):145–152. [PubMed]
  • Moreau S, Perly B, Biguet J. Interactions de la chloroquine avcc la ferriprotoporphyrine IX. Etude par résonance magnétique nucléaire. Biochimie. 1982 Nov-Dec;64(11-12):1015–1025. [PubMed]
  • Adams PA, Berman PA, Egan TJ, Marsh PJ, Silver J. The iron environment in heme and heme-antimalarial complexes of pharmacological interest. J Inorg Biochem. 1996 Jul;63(1):69–77. [PubMed]
  • Chou AC, Chevli R, Fitch CD. Ferriprotoporphyrin IX fulfills the criteria for identification as the chloroquine receptor of malaria parasites. Biochemistry. 1980 Apr 15;19(8):1543–1549. [PubMed]
  • Geary TG, Divo AD, Jensen JB, Zangwill M, Ginsburg H. Kinetic modelling of the response of Plasmodium falciparum to chloroquine and its experimental testing in vitro. Implications for mechanism of action of and resistance to the drug. Biochem Pharmacol. 1990 Aug 15;40(4):685–691. [PubMed]
  • Warhurst DC. The quinine-haemin interaction and its relationship to antimalarial activity. Biochem Pharmacol. 1981 Dec 15;30(24):3323–3327. [PubMed]
  • Ginsburg H, Demel RA. Interactions of hemin, antimalarial drugs and hemin-antimalarial complexes with phospholipid monolayers. Chem Phys Lipids. 1984 Oct;35(4):331–347. [PubMed]
  • Sugioka Y, Suzuki M, Sugioka K, Nakano M. A ferriprotoporphyrin IX-chloroquine complex promotes membrane phospholipid peroxidation. A possible mechanism for antimalarial action. FEBS Lett. 1987 Nov 2;223(2):251–254. [PubMed]
  • Mueller S, Riedel HD, Stremmel W. Direct evidence for catalase as the predominant H2O2 -removing enzyme in human erythrocytes. Blood. 1997 Dec 15;90(12):4973–4978. [PubMed]
  • Awasthi YC, Beutler E, Srivastava SK. Purification and properties of human erythrocyte glutathione peroxidase. J Biol Chem. 1975 Jul 10;250(13):5144–5149. [PubMed]
  • Ben-Yoseph Y, Shapira E. Specific immunoassay for quantitative determination of human erythrocyte catalase. J Lab Clin Med. 1973 Jan;81(1):133–139. [PubMed]
  • Atamna H, Ginsburg H. Origin of reactive oxygen species in erythrocytes infected with Plasmodium falciparum. Mol Biochem Parasitol. 1993 Oct;61(2):231–241. [PubMed]
  • Fitch CD, Yunis NG, Chevli R, Gonzalez Y. High-affinity accumulation of chloroquine by mouse erythrocytes infected with Plasmodium berghei. J Clin Invest. 1974 Jul;54(1):24–33. [PMC free article] [PubMed]
  • Martin SK, Oduola AM, Milhous WK. Reversal of chloroquine resistance in Plasmodium falciparum by verapamil. Science. 1987 Feb 20;235(4791):899–901. [PubMed]
  • Ohsawa K, Tanabe K, Kimata I, Miki A. Ultrastructural changes associated with reversal of chloroquine resistance by verapamil in Plasmodium chabaudi. Parasitology. 1991 Oct;103(Pt 2):185–189. [PubMed]
  • Kyle DE, Milhous WK, Rossan RN. Reversal of Plasmodium falciparum resistance to chloroquine in Panamanian Aotus monkeys. Am J Trop Med Hyg. 1993 Jan;48(1):126–133. [PubMed]
  • Martiney JA, Cerami A, Slater AF. Verapamil reversal of chloroquine resistance in the malaria parasite Plasmodium falciparum is specific for resistant parasites and independent of the weak base effect. J Biol Chem. 1995 Sep 22;270(38):22393–22398. [PubMed]
  • Ward SA, Bray PG, Hawley SR. Quinoline resistance mechanisms in Plasmodium falciparum: the debate goes on. Parasitology. 1997;114 (Suppl):S125–S136. [PubMed]
  • Kelder PP, Fischer MJ, de Mol NJ, Janssen LH. Oxidation of chlorpromazine by methemoglobin in the presence of hydrogen peroxide. Formation of chlorpromazine radical cation and its covalent binding to methemoglobin. Arch Biochem Biophys. 1991 Feb 1;284(2):313–319. [PubMed]
  • Bitonti AJ, Sjoerdsma A, McCann PP, Kyle DE, Oduola AM, Rossan RN, Milhous WK, Davidson DE., Jr Reversal of chloroquine resistance in malaria parasite Plasmodium falciparum by desipramine. Science. 1988 Dec 2;242(4883):1301–1303. [PubMed]
  • Bray PG, Mungthin M, Ridley RG, Ward SA. Access to hematin: the basis of chloroquine resistance. Mol Pharmacol. 1998 Jul;54(1):170–179. [PubMed]
  • Sanchez CP, Wünsch S, Lanzer M. Identification of a chloroquine importer in Plasmodium falciparum. Differences in import kinetics are genetically linked with the chloroquine-resistant phenotype. J Biol Chem. 1997 Jan 31;272(5):2652–2658. [PubMed]
  • Malhotra K, Salmon D, Le Bras J, Vilde JL. Potentiation of chloroquine activity against Plasmodium falciparum by the peroxidase-hydrogen peroxide system. Antimicrob Agents Chemother. 1990 Oct;34(10):1981–1985. [PMC free article] [PubMed]
  • Dubois VL, Platel DF, Pauly G, Tribouley-Duret J. Plasmodium berghei: implication of intracellular glutathione and its related enzyme in chloroquine resistance in vivo. Exp Parasitol. 1995 Aug;81(1):117–124. [PubMed]
  • Macomber PB, Sprinz H. Morphological effects of chloroquine on Plasmodium berghei in mice. Nature. 1967 May 27;214(5091):937–939. [PubMed]
  • el-Shoura SM. Falciparum malaria in naturally infected human patients: VIII. Fine structure of intraerythrocytic asexual forms before and during chloroquine treatment. Appl Parasitol. 1994 Sep;35(3):207–218. [PubMed]

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