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Purification, characterization, structural analysis and protein chemistry of a buffalo heart galectin-1

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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.

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Acknowledgments

The financial assistance in the form of scholarship to the authors from University Grants Commission (UGC), New Delhi is thankfully acknowledged. The authors are grateful to Aligarh Muslim University, UGC-SAP and DST-FIST for the lab facilities. The assistance provided by Dr. Syed Mohd. Kashif Rais Zaidi (Stanford University, USA) and Mr. Ausaf Ahmad (Central Drug Research Institute, Lucknow) is gratefully acknowledged.

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  1. Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, Uttar Pradesh, India

    Ghulam Md Ashraf, Sabika Rizvi, Shoa Naqvi, Nida Suhail, Nayeem Bilal, Shirin Hasan, Mohammad Tabish & Naheed Banu

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  1. Ghulam Md Ashraf
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  2. Sabika Rizvi
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  3. Shoa Naqvi
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  7. Mohammad Tabish
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  8. Naheed Banu
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Correspondence to Naheed Banu.

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Ashraf, G.M., Rizvi, S., Naqvi, S. et al. Purification, characterization, structural analysis and protein chemistry of a buffalo heart galectin-1. Amino Acids 39, 1321–1332 (2010). https://doi.org/10.1007/s00726-010-0574-7

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  • DOI: https://doi.org/10.1007/s00726-010-0574-7

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