Interactions of pig brain cytosolic sialidase with gangliosides. the formation of catalytically inactive enzyme-ganglioside complexes requires. homogeneous ganglioside micelles and is a reversible phenomenon

Bruno Venerando, Amelia Fiorilli, Luigi Caimi, Guido Tettamanti

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2 Citations (Scopus)

Abstract

Cytosolic sialidase A, obtained from pig brain and purified, interacts with ganglioside GT1b giving two catalytically inactive enzyme-ganglioside complexes. Treatment of these complexes with Triton X-100 under given conditions (1% detergent; 1 h at 37°C; 0.1 M acetic acid-sodium acetate buffer, pH4.8) leads to the liberation of part of the enzyme (about 47%) in a free and fully active form. Reversible inactivation of cytosolic sialidase requires the presence of homogeneous micelles of GTlb or of mixed micelles (for instance Triton X-100 and GT1b) with a high GT1b content. Triton X-100/ganglioside mixed micelles with a molar ratio above 50, as well as small unilamellar vesicles of egg yolk lecithin and GTlb (7-15 mol%), did not inactivate the enzyme at all; on the contrary these forms of ganglioside dispersion behaved as excellent substrates for the enzyme. It is to be concluded that under in vitro conditions the ability of ganglioside to interact with cytosolic sialidase, giving rise to catalytically inactive complexes or to Michaelis-Menten enzyme-substrate complexes, depends on the supramolecular organization of the ganglioside molecules. Arrangements of tightly packed molecules with strong side-side interactions facilitate the formation of complexes with the enzyme; arrangement with separated and loosely interacting molecules facilitates binding at the catalytically active site of the enzyme.

Original languageEnglish
Pages (from-to)1167-1176
Number of pages10
JournalJournal of Biochemistry
Volume102
Issue number5
Publication statusPublished - Nov 1987

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Micelles
Gangliosides
Neuraminidase
Brain
Enzymes
Swine
Interaction
Octoxynol
Molecules
Arrangement
Substrate
Lecithin
Sodium Acetate
Unilamellar Liposomes
Egg Yolk
Lecithins
Detergents
Vesicles
Substrates
Pig

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Physiology (medical)
  • Radiology Nuclear Medicine and imaging
  • Molecular Biology
  • Biochemistry

Cite this

Interactions of pig brain cytosolic sialidase with gangliosides. the formation of catalytically inactive enzyme-ganglioside complexes requires. homogeneous ganglioside micelles and is a reversible phenomenon. / Venerando, Bruno; Fiorilli, Amelia; Caimi, Luigi; Tettamanti, Guido.

In: Journal of Biochemistry, Vol. 102, No. 5, 11.1987, p. 1167-1176.

Research output: Contribution to journalArticle

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abstract = "Cytosolic sialidase A, obtained from pig brain and purified, interacts with ganglioside GT1b giving two catalytically inactive enzyme-ganglioside complexes. Treatment of these complexes with Triton X-100 under given conditions (1{\%} detergent; 1 h at 37°C; 0.1 M acetic acid-sodium acetate buffer, pH4.8) leads to the liberation of part of the enzyme (about 47{\%}) in a free and fully active form. Reversible inactivation of cytosolic sialidase requires the presence of homogeneous micelles of GTlb or of mixed micelles (for instance Triton X-100 and GT1b) with a high GT1b content. Triton X-100/ganglioside mixed micelles with a molar ratio above 50, as well as small unilamellar vesicles of egg yolk lecithin and GTlb (7-15 mol{\%}), did not inactivate the enzyme at all; on the contrary these forms of ganglioside dispersion behaved as excellent substrates for the enzyme. It is to be concluded that under in vitro conditions the ability of ganglioside to interact with cytosolic sialidase, giving rise to catalytically inactive complexes or to Michaelis-Menten enzyme-substrate complexes, depends on the supramolecular organization of the ganglioside molecules. Arrangements of tightly packed molecules with strong side-side interactions facilitate the formation of complexes with the enzyme; arrangement with separated and loosely interacting molecules facilitates binding at the catalytically active site of the enzyme.",
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