Paracoccin: Purification and Validation of Its Lectin and Enzymatic Properties

Methods Mol Biol. 2020:2132:139-149. doi: 10.1007/978-1-0716-0430-4_14.

Abstract

Studies on the effects of components derived from the human pathogenic fungi Paracoccidioides brasiliensis have identified paracoccin (PCN), as a bifunctional protein with lectin (GlcNAc-binding) and enzymatic (chitinase) activities, able to induce modulation of host immune response. Endogenous PCN acts as a fungal virulence factor, whereas exogenous purified PCN, administered to the host, confers protective immunity in a murine model of paracoccidioidomycosis. The immunomodulation induced by purified-PCN injection has characterized it as an agent applicable in the therapy and vaccine against paracoccidioidomycosis. This section describes methods for PCN purification and validation of its lectin and enzymatic activities. It includes detailed protocols to obtain homogeneous PCN from P. brasiliensis yeasts, as well as to purify recombinant PCN from transformed heterologous microorganisms.

Keywords: Biological properties; Chitinase; Lectin purification; P. brasiliensis; Paracoccin.

MeSH terms

  • Acetylglucosamine / metabolism*
  • Animals
  • Chitinases / metabolism
  • Disease Models, Animal
  • Fungal Proteins / administration & dosage*
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism
  • Lectins / administration & dosage*
  • Lectins / genetics
  • Lectins / isolation & purification
  • Lectins / metabolism
  • Mice
  • Paracoccidioides / immunology
  • Paracoccidioides / metabolism
  • Paracoccidioides / pathogenicity*
  • Paracoccidioidomycosis / immunology
  • Paracoccidioidomycosis / prevention & control*
  • Protein Binding
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / metabolism
  • Virulence Factors / genetics
  • Virulence Factors / metabolism

Substances

  • Fungal Proteins
  • Lectins
  • Recombinant Proteins
  • Virulence Factors
  • paracoccin, Paracoccidioides brasiliensis
  • Chitinases
  • Acetylglucosamine