Isolation and some properties of a glycoprotein of 70 kDa (Gp70) from the cell wall of Sporothrix schenckii involved in fungal adherence to dermal extracellular matrix.
Sporothrix schenckii is the etiological agent of sporotrichosis, a subcutaneous mycosis and an emerging disease in immunocompromised patients. Adherence to target cells is a prerequisite for fungal dissemination and systemic complications. However, information on the cell surface components involved in this interaction is rather scarce. In this investigation, the extraction of isolated cell walls from the yeast phase of S. schenckii with SDS and separation of proteins by SDS-PAGE led to the identification of a periodic acid-Schiff (PAS)-reacting 70 kDa glycoprotein (Gp70) that was purified by elution from electrophoresis gels. The purified glycopeptide exhibited a pI of 4.1 and about 5.7% of its molecular mass was contributed by N-linked glycans with no evidence for O-linked oligosaccharides. Confocal analysis of immunofluorescence assays with polyclonal antibodies directed towards Gp70 revealed a rather uniform distribution of the antigen at the cell surface with no distinguishable differences among three different isolates. Localization of Gp70 at the cell surface was confirmed by immunogold staining. Gp70 seems specific for S. schenckii as no immunoreaction was observed in SDS-extracts from other pathogenic and non-pathogenic fungi. Yeast cells of the fungus abundantly adhered to the dermis of mouse tails and the anti-Gp70 serum reduced this process in a concentration-dependent manner. Results are discussed in terms of the potential role of Gp70 in the host-pathogen interaction.
Ruiz-Baca
Estela
E
Instituto de Investigación en Biología Experimental, Facultad de Química, Universidad de Guanajuato, Guanajuato, México.
Toriello
Conchita
C
Perez-Torres
Armando
A
Sabanero-Lopez
Myrna
M
Villagomez-Castro
Julio C
JC
Lopez-Romero
Everardo
E
D000818:Animals; D002448:Cell Adhesion; D002473:Cell Wall / Q000378:metabolism*; D020405:Dermis / Q000382:microbiology* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000382:microbiology*; D006023:Glycoproteins / Q000302:isolation & purification / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D051379:Mice; D011817:Rabbits; D013175:Sporothrix / Q000378:metabolism / Q000502:physiology*; D013174:Sporotrichosis / Q000382:microbiology*
D006023:Glycoproteins