Specific Staining of Wall Mannan in Yeast Cells with Fluorescein-Conjugated Concanavalin A

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Abstract
A procedure is given for the coupling of fluorescein isothiocyanate to concanavalin A, a protein which specifically combines with a variety of polysaccharides, and for the subsequent isolation of the reactive conjugate. This fluorescent conjugate stains Saccharomyces cerevisiae but not Schizosaccharomyces pombe or Rhodotorula glutinis . The cell walls of the latter two organisms do not contain branched homopolymers of α-linked mannose. Furthermore, the staining of S. cerevisiae is competitively inhibited by either unlabeled concanavalin A or methyl-α- d -manno-pyranoside. On the basis of this evidence, it is concluded that the staining of S. cerevisiae results from the specific interaction of the fluorescein-concanavalin A conjugate with the α-mannan present in the cell wall of this yeast.

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