Radiolabeling of lectin from Robinia pseudoacacia by reductive methylation.
Lectin from the bark of Robinia pseudoacacia was radiolabeled by reductive methylation. Reductive methylation of the lectin with [14C]formaldehyde and a reducing agent, sodium borohydride, resulted in incorporation of 14C into the lectin but with considerable inactivation of carbohydrate binding activity. Dialysis of the labeled lectin against the buffer containing Ca2+ and Mn2+ partially restored the activity. Higher incorporation of 14C into the lectin was observed when sodium cyanoborohydride was used as a reducing agent. In this case, more than eighty percent of the labeled lectin retained carbohydrate binding activity. Autoradiography showed that the radiolabeled lectin had a molecular mass of 29 kDa, corresponding to that of the intact lectin subunit.
Tazaki
K
K
Forestry and Forest Products Research Institute, Ibaraki, Japan.
D002250:Carbon Radioisotopes; D007553:Isotope Labeling; D037102:Lectins / Q000737:chemistry*; D008745:Methylation; D010084:Oxidation-Reduction; D037121:Plant Lectins
D002250:Carbon Radioisotopes; D037102:Lectins; D037121:Plant Lectins; C047720:Robinia lectin