Kluyveromyces bulgaricus yeast lectins. Isolation of two galactose-specific lectin forms from the yeast cell wall.
Incubation of galactose treated Kluyveromyces bulgaricus yeast cells in EDTA/phosphate-buffered saline led to an extract possessing hemagglutinating and yeast flocculating properties. Purification of this extract by affinity chromatography and gel filtration gave two lectin forms, Kb-CWL I and Kb-CWL II, with an apparent molecular mass of 38,000 and 150,000 Da, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that Kb-CWL I and Kb-CWL II were dimeric and octameric of a subunit of 18,900 Da. At high concentration, purified Kb-CWL I associated to give Kb-CWL II. This association seemed to be independent on pH. The two lectin forms were glycoproteins, the peptide counterpart was very rich in Lys, Glu, and Gly, and the carbohydrate part represented 1% of the whole molecule and was composed of Glc, Man, and Ara. The two lectin forms (KB-CWL I and Kb-CWL II) agglutinated human red blood cells and flocculated EDTA-treated K. bulgaricus yeast cells. The activity of both lectin forms required Ca2+ ions, while Sr2+ showed some competitive inhibition. Optimal activity was obtained within a pH range of 4-6.5 for both forms. Temperatures of 80-90 degrees C for 20 min, or proteolytic treatment reduced irreversibly the activity of Kb-CWL I and Kb-CWL II. The role of the cell wall phosphopeptidomannan as a ligand and a potential physiological receptor of these lectin forms was demonstrated.
Laboratoire de Biochimie Microbienne, Faculté des Sciences Pharmaceutiques et Biologiques, Nancy, France.
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al-Mahmood
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Colin
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Bonaly
D000371:Agglutination; D000596:Amino Acids / Q000032:analysis; D001667:Binding, Competitive; D002412:Cations; D002473:Cell Wall / Q000737:chemistry; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D037161:Galectins; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006388:Hemagglutinins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D007716:Kluyveromyces / Q000737:chemistry*; D008024:Ligands; D008351:Mannans / Q000378:metabolism*; D008970:Molecular Weight
D000596:Amino Acids; D002412:Cations; D005656:Fungal Proteins; D037161:Galectins; D006023:Glycoproteins; D006388:Hemagglutinins; D008024:Ligands; D008351:Mannans