The fungal wall protects cells
against extracellular challenges, and allows them to interact with the
surroundings. A major component of fungal cell walls is the
polysaccharide beta-1,3-glucan. Synthesis of beta-1,3-glucan has been
quantified in two ways. The first method makes use of radioactive
UDP-glucose, and the amount of in vitro produced
beta-1,3-glucan that can be precipitated with trichloric acid
(Shematek et al., 1980. J. Biol. Chem. 255:
888-94). The other method utilizes a fluorescent dye that specifically
stains the in vitro synthesized beta-1,3-glucan, and the amount
of which can be quantified in a fluorescent plate reader (Shedletzky
et al., 1997. Anal. Biochem. 249: 88-93). Both
methods require expensive materials and/or equipment for assaying the
beta-1,3-glucan activity. Here we use a competitive Enzyme
Immunoassay (Douwes et al.,
1996. Appl. Environ. Microbiol. 62: 3176-82) for
measuring product formation in an in vitro beta-1,3-glucan
synthase assay. This method can be used routinely for large-scale
sample analysis. With the availability of commercial monoclonal
antibodies this method provides a reliable and relatively low-cost
quantification of the activity of beta-1,3-glucan synthase.
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