Purification and some properties of galectin-1 derived from water buffalo (Bubalus bubalis) brain.
An increasing number of galectins have been found in various animal species, the most abundant of which is galectin-1. The purpose of the present study was to purify and characterize galectin-1 from buffalo brain. We purified the galectin using a combination of ammonium sulphate fractionation and affinity chromatography and the homogeneity was determined by both native polyacrylamide gel electrophoresis (PAGE) and denaturing SDS-PAGE. The molecular weight of the galectin as determined by SDS-PAGE under reducing conditions and by gel filtration column under native conditions was 13.8 and 24.5 kDa, respectively, suggesting a dimeric form of galectin. The most potent inhibitor of the galectin activity was lactose, giving complete inhibition of hemagglutination at 0.8 mM. Galectin showed higher specificity towards human blood group A. Free thiol groups were estimated at a molar ratio of 2.9. The effects of alkylating reagents (iodoacetate and iodoacetamide) on saccharide binding of the galectin were studied. Both alkylating reagents significantly inactivated the activity of the galectin within 20 min. The temperature and pH stability of the galectin were determined. Our findings based on physico-chemical properties, carbohydrate and blood group specificities of the galectin may have future implications in biological and clinical applications.
Ola
M Shamsul
MS
Department of Biochemistry, Faculty of Life Sciences, A.M.U. Aligarh, UP, India. sola@psu.edu
Tabish
M
M
Khan
F H
FH
Banu
Naheed
N
D000017:ABO Blood-Group System; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002020:Buffaloes / Q000378:metabolism*; D050260:Carbohydrate Metabolism / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D037483:Galectin 1 / Q000302:isolation & purification* / Q000378:metabolism*; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007460:Iodoacetamide / Q000494:pharmacology; D007461:Iodoacetates / Q000494:pharmacology; D008970:Molecular Weight; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013696:Temperature
D000017:ABO Blood-Group System; D037483:Galectin 1; D007461:Iodoacetates; D013438:Sulfhydryl Compounds; D007460:Iodoacetamide