TY - JOUR
T1 - Purification, characterization, structural analysis and protein chemistry of a buffalo heart galectin-1
AU - Ashraf, Ghulam Md
AU - Rizvi, Sabika
AU - Naqvi, Shoa
AU - Suhail, Nida
AU - Bilal, Nayeem
AU - Hasan, Shirin
AU - Tabish, Mohammad
AU - Banu, Naheed
PY - 2010/11
Y1 - 2010/11
N2 - 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.
AB - 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.
KW - 3D-JIGSAW
KW - Bubalus bubalis
KW - Buffalo heart galectin-1 (BHG-1)
KW - Cardiovascular disorders
KW - Oxidation
KW - Purification
KW - Sequencing
UR - http://www.scopus.com/inward/record.url?scp=78449269803&partnerID=8YFLogxK
U2 - 10.1007/s00726-010-0574-7
DO - 10.1007/s00726-010-0574-7
M3 - Article
C2 - 20354738
AN - SCOPUS:78449269803
SN - 0939-4451
VL - 39
SP - 1321
EP - 1332
JO - Amino Acids
JF - Amino Acids
IS - 5
ER -