Identification of hemoglobin degradation products in Plasmodium falciparum.
Malaria parasites break down human hemoglobin to its constituent amino acids by cysteine and aspartic proteinases. However, no one has previously been able to identify hemoglobin cleavage products in intact parasites. When isolated parasites were subjected to non-denaturing polyacrylamide gels electrophoresis, a unique protein band was found which contains heme and reacts with anti-human hemoglobin antibodies. This protein does not appear to represent oxidized or glycosylated hemoglobin, and is present in isolated parasites but not in the cytosol of infected or uninfected erythrocytes. When this band was eluted and subjected to SDS polyacrylamide gel electrophoresis, three bands were seen on Western blots. The proteins in these bands contain proteins with the N-terminal sequences of alpha- and beta-globin chains but molecular masses of only 13.2-13.4 kDa. These data suggest that hemoglobin alpha- and beta-chains are initially cleaved within the parasite phagolysosome to release peptides of 15-17 and 23-25 amino acids from the C-termini of alpha- and beta-globin chains, respectively. Production of the hemoglobin breakdown products was inhibited by E-64, a cysteine proteinase inhibitor, suggesting the involvement of a cysteine proteinase in an early step of hemoglobin degradation.
Kamchonwongpaisan
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Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor 48109-2029, USA.
Samoff
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E
Meshnick
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SR
D000595:Amino Acid Sequence; D000818:Animals; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000032:analysis / Q000187:drug effects; D005914:Globins / Q000378:metabolism; D006454:Hemoglobins / Q000737:chemistry / Q000378:metabolism*; D015151:Immunoblotting; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010436:Pepstatins / Q000494:pharmacology; D010963:Plasmodium falciparum / Q000737:chemistry / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D015800:Protozoan Proteins / Q000737:chemistry
D015853:Cysteine Proteinase Inhibitors; D006454:Hemoglobins; D010436:Pepstatins; D011480:Protease Inhibitors; D015800:Protozoan Proteins; C014195:Streptomyces pepsin inhibitor; D005914:Globins; D010450:Endopeptidases; D007930:Leucine; C024974:E 64; C031375:pepstatin