Kinetics of yeast dissociation from lectin beads: I. alpha methyl mannose

Abstract

Here we examine the kinetics of alpha methyl mannose induced dissociation of yeast (Saccharomyces cerevisiae) that have been previously allowed to bind to concanavalin A (Con A) derivatized agarose beads, a model for eluting glycans from Con A beads in purification protocols and for releasing pathogens from human cells. This study tests the statistical reliability of this model using one glycan concentration with hundreds of replicate experiments. Future dose‐response studies will precisely identify the reagents that are most effective at the lowest concentrations. In 963 trials, yeast release from the Con A beads was measured by counting yeast remaining on single yeast bound beads in the presence and absence of 1.5M alpha methyl mannose over a 60 min time course. Two‐ sample t tests showed significant differences in the amount of yeast remaining bound in the sugar versus control (no sugar) samples after 20 min (p less than 0.0001), 40 min (p less than 0.0001) and 60 min (p less than 0.0001), where the sugar caused substantially more dissociation than in its absence. Repeated measures‐ANOVA indicated a significant effect of the glycan treatment on the number of bound yeast (F= 624, p less than 0.0001), as well as a significant interaction between the treatment and time elapsed (F= 339, p less than 0.0001). This is therefore a reliable, simple, precise and inexpensive model for dissociation kinetic studies. Two other studies in these proceeding using different reagents support this conclusion (Supported by NIH NIGMS SCORE (S0648680), RISE, MARC, the Joseph Drown Foundation and the Sidney Stern Memorial Trust).