Comparative rates of sialylation by recombinant trans-sialidase and inhibitor properties of synthetic oligosaccharides from Trypanosoma cruzi mucins-containing galactofuranose and galactopyranose.
The mucin-like glycoproteins of Trypanosoma cruzi have novel O-linked oligosaccharides that are acceptors of sialic acid in the trans-sialidase (TcTS) reaction. The transference of sialic acid from host glycoconjugates to the mucins is involved in infection and pathogenesis. The O-linked chains may contain galactofuranose in addition to the acceptor galactopyranose units. Thus far, the galactofuranose form was found in the mucins of strains belonging to the less infective lineage. The acceptor properties of the chemically synthesized oligosaccharides were now studied in order to correlate their structure with the ability to act as substrates. Recombinant TcTS and sialyllactose as donor were used. The reactions were followed by HPAEC-PAD. The K(m) values were calculated for the free sugars, the sugar alditols and the benzyl glycosides. All the compounds showed to be good acceptors of sialic acid. Thus, the introduction of galactofuranose in the mucins of the strains of lineage 1 would not be responsible for the diminished virulence of the strains. The oligosaccharides and derivatives inhibited the transfer of sialic acid to the substrate N-acetyllactosamine with IC(50) values between 0.6 and 4 mM.
Agustí
Rosalía
R
CIHIDECAR, Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pabellón II, 1428 Buenos Aires, Argentina.
Giorgi
M Eugenia
ME
Mendoza
Verónica M
VM
Gallo-Rodriguez
Carola
C
de Lederkremer
Rosa M
RM
D000606:Amino Sugars / Q000737:chemistry; D000818:Animals; D002240:Carbohydrate Sequence; D004926:Escherichia coli / Q000378:metabolism; D005690:Galactose / Q000737:chemistry* / Q000302:isolation & purification; D007700:Kinetics; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry*; D009439:Neuraminidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000737:chemistry; D012794:Sialic Acids / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000737:chemistry*
D000606:Amino Sugars; D009077:Mucins; D009844:Oligosaccharides; D011994:Recombinant Proteins; D012794:Sialic Acids; C000458:N-acetyllactosamine; D009439:Neuraminidase; D005690:Galactose