Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor.
Here we report the presence of a trans-sialidase on the surface of Trypanosoma brucei culture-derived procyclic trypomastigotes. The enzyme is not detected in lysates of bloodstream trypomastigotes enriched for either stumpy or slender forms. The trans-sialidase catalyzes the transfer of alpha(2-3)-linked sialic acid residues to lactose. beta-galactopyranosyl residues are at least 100 times better acceptors for sialic acid than alpha-galactopyranosyl residues. In the absence of efficient acceptors, the purified enzyme transfers sialic acid to water, i.e., it acts as a sialidase. Although the T. cruzi and T. brucei trans-sialidases have very similar donor and acceptor specificities, they are antigenically distinct. Sodium dodecyl sulfate-polyacramide gel electrophoresis under nonreducing conditions and silver staining of the purified trans-sialidase reveals a single band of 63 kD. When the surface membrane of live procyclic trypomastigotes is trans-sialylated, using radioactive sialyllactose as the donor substrate, it appears that the only sialylated surface molecule is procyclin. Pronase treatment of live parasites removes only part of the surface sialic acid, in agreement with recent data showing that the glycosylphosphatidylinositol anchor of procyclin is sialylated (Ferguson, M. A. J., M. Murray, H. Rutherford, and M. J. McConville. 1993. Biochem. J. In press).
Pontes de Carvalho
L C
LC
Department of Pathology, New York University Medical Center, New York 10016.
Tomlinson
S
S
Vandekerckhove
F
F
Bienen
E J
EJ
Clarkson
A B
AB
Jiang
M S
MS
Hart
G W
GW
Nussenzweig
V
V
D000818:Animals; D008562:Membrane Glycoproteins; D009439:Neuraminidase / Q000378:metabolism; D015800:Protozoan Proteins / Q000302:isolation & purification / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism; D012795:Sialoglycoproteins / Q000378:metabolism; D012799:Sialyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000166:cytology / Q000201:enzymology*; D014349:Trypanosoma cruzi / Q000201:enzymology; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000378:metabolism*
D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D012794:Sialic Acids; D012795:Sialoglycoproteins; D014643:Variant Surface Glycoproteins, Trypanosoma; C053277:procyclic acidic repetitive protein, Trypanosoma; D012799:Sialyltransferases; D009439:Neuraminidase