A single tyrosine differentiates active and inactive Trypanosoma cruzi trans-sialidases.
Several genes encode members of the Trypanosoma cruzi (Tc) trans-sialidase (TS) family. These proteins contain an enzymatic domain on the N terminus, the only one required for TS activity, and an antigenic domain (SAPA (shed acute phase antigen) amino acid (aa) repeats) on the C terminus. Only some members of this glycoprotein family are enzymatically active. The complete sequence of two clones encoding the enzymatic domain of active and inactive protein from each of two Tc strains has now been obtained. Comparison of these sequences showed a limited divergence among them: 20 out of the 642 deduced aa in the enzymatic domain were found to differ. From these 20 aa, only one was found to be essential for enzymatic activity. A Tyr342 residue is deduced in both active proteins while a His342 is present in both inactive ones. This naturally occurring Tyr342-->His substitution completely abolished the TS activity. In addition to Tyr342, a second deduced aa, Pro231, was found to be necessary for full enzymatic TS activity; a Pro231-->Ala change rendered the TS protein partially active. Fourteen aa residues, including Tyr342, out of the 16 aa in the active site of a sialidase from Salmonella typhimurium are present at the same or very similar positions in the Tc TS.
Cremona
M L
ML
Instituto de Investigaciones Bioquímicas Fundación Campomar, Buenos Aires, Argentina.
Sánchez
D O
DO
Frasch
A C
AC
Campetella
O
O
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D017125:Genes, Protozoan; D014644:Genetic Variation; D007700:Kinetics; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D013045:Species Specificity; D014349:Trypanosoma cruzi / Q000201:enzymology* / Q000235:genetics; D014443:Tyrosine
D017931:DNA Primers; D014443:Tyrosine; D009439:Neuraminidase