Purification and partial characterization of a 32-kilodalton sialic acid-specific lectin from Aspergillus fumigatus.
Adherence of the opportunistic fungus Aspergillus fumigatus to the extracellular matrix components is considered a crucial step in the establishment of the infection. Given the high carbohydrate content of these glycoproteins and the role of carbohydrate-protein interactions in numerous adherence processes, the presence of a lectin in A. fumigatus was investigated. Different fungal extracts obtained by sonication or grinding in liquid nitrogen from resting or swollen conidia, as well as from germ tubes and mycelium, were tested by hemagglutination assays using rabbit erythrocytes. A lectin activity was recovered in all the extracts tested. However, sonication of resting conidia resulted in the highest specific activity. Purification of the lectin was achieved by gel filtration followed by ion-exchange and hydrophobic-interaction chromatographies. Analysis of the purified lectin by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed an apparent molecular mass of 32 kDa, which is similar to that of the alkaline protease already identified from different strains of A. fumigatus. However, as evidenced by the use of an alkaline protease-deficient mutant, the two activities were supported by distinct proteins. In addition, hemagglutination inhibition experiments using different saccharides and glycoproteins demonstrated the specificity of the lectin for sialic acid residues. Together these results suggest that this lectin may contribute to the attachment of conidia to the extracellular matrix components through the recognition of the numerous terminal sialic acid residues of their carbohydrate chains.
Tronchin
Guy
G
Groupe d'Etude des Interactions Hôte-Parasite, UPRES EA 3142, Laboratoire de Parasitologie-Mycologie, Centre Hospitalier Universitaire, Angers, France. guy.tronchin@univ-angers.fr
Esnault
Karine
K
Sanchez
Myriam
M
Larcher
Gerald
G
Marot-Leblond
Agnes
A
Bouchara
Jean-Philippe
JP
D000818:Animals; D001232:Aspergillus fumigatus / Q000235:genetics / Q000378:metabolism*; D006001:Glycoconjugates / Q000737:chemistry / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests; D006386:Hemagglutination Tests; D037102:Lectins / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D011817:Rabbits; D013379:Substrate Specificity
D006001:Glycoconjugates; D037102:Lectins; D019158:N-Acetylneuraminic Acid