Solubilization of soybean membrane binding sites for fungal β‐glucans that elicit phytoalexin accumulation

Soybean membranes contain high-affinity binding sites for fungal beta-glucans. These sites may play a role in the recognition by soybean tissues of fungal phytoalexin elicitors. We have solubilized beta-glucan-binding activity from microsomal membranes using two C12-alkyl zwitterionic detergents, Zwittergent 3-12 (ZW 3-12) and the lysolecithin analog 1-dodecanoyl propanediol-3-phosphorylcholine [corrected] (ES12H). The solubilized binding sites displayed identical affinity for beta-glucans as that found in membranes (KD = 11-34 nM). Detergent-protein micelles with glucan binding activity eluted with approximate Mr values of 300,000 in ZW 3-12 and 380,000 in ES12H in gel permeation chromatography. Maximal binding activity eluted from a chromatofocusing column in the pH range between 6.2 and 6.6 with both ES12H and ZW 3-12, suggesting an apparent pI close to neutral.