Isolation and characterization of beta-galactoside specific lectin from Korean mistletoe (Viscum album var. coloratum) with lactose-BSA-sepharose 4B and changes of lectin conformation.
Lectins and its A- and B-chains from Korean mistletoe (Viscum album var. coloratum) were isolated by affinity chromatography on the Sepharose 4B modified by lactose-BSA conjugate synthesized by reductive amination of ligand (lactose) to epsilon-amino groups of lysine residues of spacer (BSA) after reduction by NaCNBH3. The lactose-BSA conjugate was coupled to Sepharose 4B activated by cyanogen bromide. The molecular weight determined by SDS-PAGE were a 31 kD of A-chain and a 35 kD of B-chain. Amino acid analysis and N-terminal sequencing were performed. The effects of pH, temperature and guanidine chloride on the conformation of the lectin were investigated by measuring its intrinsic fluorescence and compared with its hemagglutinating activities. Blue shift was detected on the acidic pH and there was a close relationship between activities and conformation of the lectin. Under denaturing conditions, the tryptophan emission profile of lectin showed typical denaturational red shift which also correspond to the conformations and activity of lectin.
Park
W B
WB
Faculty of Environmental and Life Science, Seoul Women's University, Korea.
Ju
Y J
YJ
Han
S K
SK
D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005697:Galactosides / Q000378:metabolism*; D019791:Guanidine / Q000494:pharmacology; D006386:Hemagglutination Tests; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000737:chemistry* / Q000302:isolation & purification*; D014784:Mistletoe / Q000737:chemistry*; D037121:Plant Lectins; D010946:Plants, Medicinal; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation / Q000187:drug effects; D013379:Substrate Specificity; D013696:Temperature
D000596:Amino Acids; D005697:Galactosides; D037102:Lectins; D037121:Plant Lectins; C007816:beta-galactoside; D019791:Guanidine