Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor

Lain C. Pontes De Carvalho, Stephen Tomlinson, Filip Vandekerckhove, E. Jay Bienen, Allen B. Clarkson, Man Shiow Jiang, Gerald Warren Hart, Victor Nussenzweig

Research output: Contribution to journalArticle

Abstract

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 α(2-3)-linked sialic acid residues to lactose. β-galactopyranosyl residues are at least 100 times better acceptors for sialic acid than α-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.

Original languageEnglish (US)
Pages (from-to)465-474
Number of pages10
JournalJournal of Experimental Medicine
Volume177
Issue number2
StatePublished - Feb 1 1993

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Trypanosoma brucei brucei
N-Acetylneuraminic Acid
Glycosylphosphatidylinositols
Pronase
Silver Staining
Neuraminidase
Enzymes
Lactose
Sodium Dodecyl Sulfate
Electrophoresis
Parasites
Gels
Membranes
trans-sialidase
Water

ASJC Scopus subject areas

  • Immunology

Cite this

Pontes De Carvalho, L. C., Tomlinson, S., Vandekerckhove, F., Bienen, E. J., Clarkson, A. B., Jiang, M. S., ... Nussenzweig, V. (1993). Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor. Journal of Experimental Medicine, 177(2), 465-474.

Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor. / Pontes De Carvalho, Lain C.; Tomlinson, Stephen; Vandekerckhove, Filip; Bienen, E. Jay; Clarkson, Allen B.; Jiang, Man Shiow; Hart, Gerald Warren; Nussenzweig, Victor.

In: Journal of Experimental Medicine, Vol. 177, No. 2, 01.02.1993, p. 465-474.

Research output: Contribution to journalArticle

Pontes De Carvalho, LC, Tomlinson, S, Vandekerckhove, F, Bienen, EJ, Clarkson, AB, Jiang, MS, Hart, GW & Nussenzweig, V 1993, 'Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor', Journal of Experimental Medicine, vol. 177, no. 2, pp. 465-474.
Pontes De Carvalho, Lain C. ; Tomlinson, Stephen ; Vandekerckhove, Filip ; Bienen, E. Jay ; Clarkson, Allen B. ; Jiang, Man Shiow ; Hart, Gerald Warren ; Nussenzweig, Victor. / Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor. In: Journal of Experimental Medicine. 1993 ; Vol. 177, No. 2. pp. 465-474.
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