Modulation of the activity of Clostridium perfringens neuraminidase by the molecular organization of gangliosides in monolayers

María A. Perillo, Robert K. Yu, Bruno Maggio

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The activity of Clostridium perfringens neuraminidase against gangliosides GM3, GD1a and GM1 was studied in lipid monolayers at the air-buffer solution interface. The enzyme activity assay against pure ganglioside monolayers is based on the markedly different molecular packing areas of the substrate gangliosides and the resulting product glycosphingolipids. This allows to control and monitor the surface pressure and the ganglioside intermolecular organization (cross-sectional packing areas and dipole potentials) in a continuous manner during the catalytic process. It was found that the rate and the extent of the enzymatic reaction depended markedly on the lateral surface pressure. In general, the activity of neuraminidase against GM3 and GD1a was higher at lower surface pressure. This corresponded to larger intermolecular spacings among the ganglioside molecules. Both the activity and the extent of the reaction against GM3 were higher than toward GD1a. GM1 could not be degraded by the enzyme, irrespective of the surface pressure but the enzyme could interact with this ganglioside. A latency period, longer for GM3 than for GD1a, was observed prior to the onset of rapid degradation; this indicates that pre-catalytic steps are occurring at the interface before effective ganglioside degradation takes place. The latency period, the total amount of ganglioside degraded, and the velocity of the reaction varied with the surface pressure in different manners. Our data indicate that the different steps of the catalytic reaction occurring at the surface (i.e., substrate recognition and interfacial adsorption, catalysis, maximum extent of substrate conversion) are independently regulated by the molecular organization of the substrate gangliosides.

Original languageEnglish (US)
Pages (from-to)155-164
Number of pages10
JournalBBA - Biomembranes
Volume1193
Issue number1
DOIs
StatePublished - Jul 13 1994

Fingerprint

Clostridium
Clostridium perfringens
Gangliosides
Neuraminidase
Monolayers
Modulation
Pressure
Substrates
G(M3) Ganglioside
G(M1) Ganglioside
Degradation
Glycosphingolipids
Enzyme Assays
Enzyme activity
Enzymes
Catalysis
Adsorption
Assays
Buffers
Air

Keywords

  • Ganglioside
  • Monolayer
  • Neuraminidase
  • Surface hydrolysis

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Modulation of the activity of Clostridium perfringens neuraminidase by the molecular organization of gangliosides in monolayers. / Perillo, María A.; Yu, Robert K.; Maggio, Bruno.

In: BBA - Biomembranes, Vol. 1193, No. 1, 13.07.1994, p. 155-164.

Research output: Contribution to journalArticle

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