RESUMO
The malarial GBP 130 protein binds weakly to intact human erythrocytes; the binding sites seem to be located in the repeat region and this region's antibodies block the merozoite invasion. A peptide from this region (residues from 701 to 720) which binds to human erythrocytes was identified. This peptide named 2220 did not bind to sialic acid; the binding site on human erythrocyte was affected by treatment with trypsin but not by chymotrypsin. The peptide was able to inhibit Plasmodium falciparum merozoite invasion of erythrocytes. The residues F701, K703, L705, T706, E713 (FYKILTNTDPNDEVERDNAD) were found to be critical for peptide binding to erythrocytes.
Assuntos
Humanos , Animais , Eritrócitos/metabolismo , Glicoforinas/metabolismo , Proteína 1 de Superfície de Merozoito/química , Plasmodium falciparum/metabolismo , Sequência de Aminoácidos , Quimotripsina/farmacologia , Eritrócitos/efeitos dos fármacos , Eritrócitos/parasitologia , Glicoforinas/biossíntese , Proteína 1 de Superfície de Merozoito/metabolismo , Ácido N-Acetilneuramínico/metabolismo , Plasmodium falciparum/efeitos dos fármacos , Análise de Sequência de Proteína , Tripsina/farmacologiaRESUMO
We have studied the activity of a calcium dependent transglutaminase (EC 2.3.2.13) during the growth of the parasite Plasmodium falciparum inside the infected human erythrocyte. There is only one detectable transglutaminase in the two-cell-system, and its origin is erythrocytic. No activity was detected in preparations of the parasite devoid of erythrocyte cytoplasm. The Michaelis Menten constants (Km) of the enzyme for the substrates N'N'dimethlcaseine and putrescine were undistinguishable whether the cell extracts used in their determination were obtained from normal or from infected red cells. The total activity of transglutaminase in stringently synchronized cultures, measured at 0.5mM Ca2+, decreased with the maturation of the parasite. However, a fraction which became irreversibly activated and independent of calcium concentration was detected. The proportion of this fraction grew with maturation; it represented only 20 per cent of the acitivity in 20 hr-old-trophozoites while in 48-hr-schizonts it was more than 85 per cent of the total activity. The activation of this fraction of transglutaminase did not depend on an increase in the erythrocyte cytoplasmic calcium, since most of the calcium was shown to be located in the parasite.