title: Purification, characterization, structural analysis and protein chemistry of a buffalo heart galectin-1.
abstract: A soluble β-galactoside-binding lectin was purified by gel filtration chromatography from Bubalus bubalis heart. Its metal-independent nature, molecular weight of 14.5 kDa, preferential affinity for β-D-lactose, and 87-92% identity with carbohydrate recognition domain of previously reported galectin-1 confirmed its inclusion in galectin-1 subfamily. Stokes radii determination using gel filtration under reducing and non-reducing conditions revealed its homo-dimeric nature, further confirming its Gal-1 nomenclature. The purified lectin was found to be the most stable mammalian heart galectin purified till date, suggesting its preferential use in various recognition studies. Treatment of the purified lectin with oxidizing agent, thiol blocking reagents, denaturants, and detergents resulted in significant changes in UV-VIS, fluorescence, CD and FTIR spectra, which strongly emphasized the important aspect of regular secondary structure of galectins for the maintenance of their active conformation. Reduction of the activity of the purified lectin after oxidation by H2O2, with remarkable fluorescence quenching, may suggest potential role for galectin-1 in free radical-induced, oxidative stress-mediated cardiovascular disorders. The predictions of bioinformatics studies were found to be in accordance with the results obtained in wet lab.
authors.affiliation: Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh, India. ashraf.gm@gmail.com
authors.forename: Ghulam Md
authors.initials: GM
authors.lastname: Ashraf
authors.forename: Sabika
authors.initials: S
authors.lastname: Rizvi
authors.forename: Shoa
authors.initials: S
authors.lastname: Naqvi
authors.forename: Nida
authors.initials: N
authors.lastname: Suhail
authors.forename: Nayeem
authors.initials: N
authors.lastname: Bilal
authors.forename: Shirin
authors.initials: S
authors.lastname: Hasan
authors.forename: Mohammad
authors.initials: M
authors.lastname: Tabish
authors.forename: Naheed
authors.initials: N
authors.lastname: Banu
mesh_terms: D000818:Animals; D002417:Cattle; D037483:Galectin 1 / Q000097:blood / Q000737:chemistry* / Q000302:isolation & purification*; D006321:Heart; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017433:Protein Structure, Secondary; D020539:Sequence Analysis, Protein; D013696:Temperature
chemical_list: D037483:Galectin 1