The Ser-Arg-Tyr-Asp region of the major surface glycoprotein of Leishmania mimics the Arg-Gly-Asp-Ser cell attachment region of fibronectin.
The major surface glycoprotein of Leishmania, gp63, a fibronectin-like molecule, plays a key role in parasite-macrophage interaction. Binding of gp63 to macrophage receptors is inhibited by Arg-Gly-Asp-Ser (RGDS)-containing synthetic peptides of fibronectin and by antibodies to these peptides. However, gp63 lacks an RGDS tetrapeptide. We sought to identify the region of gp63 that antigenically and functionally mimics the RGDS-containing region of fibronectin. We thus synthesized on polyethylene rods overlapping tetracosapeptides covering the whole sequence of Leishmania major gp63. gp63 affinity-purified antibodies raised against fibronectin and against the RGDS-containing fibronectin decapeptide RGDSPASSKP bound specifically to gp63 residues 241-264. Subsequently, by the use of smaller peptides, the gp63 tetrapeptide 252-255 (SRYD) was identified as the minimum antibody binding segment. Single residue substitution peptide analogues showed that indeed Tyr and Gly can be alternatively substituted in the SRYD- and RGDS-containing peptides of gp63 and fibronectin, respectively, without major effects on their antibody binding capacity. Subsequently, we investigated the effect of an SRYD peptide on promastigote-macrophage interaction in vitro; treatment of macrophages with an SRYD-containing gp63 octapeptide efficiently inhibited parasite attachment to macrophage receptors. Thus, the conserved among species sequence SRYD of gp63, with significant hydrophilicity, flexibility, and beta-turn propensity features, mimics antigenically and functionally the RGDS sequence of fibronectin. We suggest that this segment constitutes the putative gp63 adhesion site.
Soteriadou
K P
KP
Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
Remoundos
M S
MS
Katsikas
M C
MC
Tzinia
A K
AK
Tsikaris
V
V
Sakarellos
C
C
Tzartos
S J
SJ
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002846:Chromatography, Affinity; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism*; D006790:Host-Parasite Interactions; D007891:Leishmania / Q000235:genetics / Q000502:physiology*; D008264:Macrophages / Q000502:physiology; D008666:Metalloendopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis / Q000138:chemical synthesis / Q000276:immunology / Q000494:pharmacology*
D000906:Antibodies; D005353:Fibronectins; D009842:Oligopeptides; C046617:arginyl-glycyl-aspartyl-serine; D008666:Metalloendopeptidases; C055146:glycoprotein gp63, Leishmania