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Purification, Characterization, Sequencing and Biological Chemistry of Galectin-1 Purified from Capra hircus (goat) Heart

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Abstract

A soluble β-galactoside binding 14.5 kDa lectin was purified from the heart of Capra hircus. Its metal independent nature, preferential affinity for β-d-lactose and 90–94% homology with carbohydrate recognition domain of previously reported galectin-1 confirmed its inclusion in galectin-1 subfamily. The secondary structures of the deduced amino acid sequences were generally conserved with previously reported Gal-1. Exposure of the purified protein to varying temperature and pH, oxidant, thiol blocking reagents, denaturants and detergents resulted in significant changes in UV (ultraviolet), fluorescence, CD (circular dichroism) and FTIR (fourier transform infra red) spectra, thus strongly emphasizing the vitality of regular secondary structure of galectins for maintaining their active conformation. Bioinformatics studies corroborated the results obtained in wet lab. Our findings based on physico-chemical properties, oxidative inactivation and structural analysis of the goat heart galectin-1 suggests significant implications in potential biological and clinical applications.

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Abbreviations

Gal-1:

Galectin-1

CD:

Circular dichroism

CBB:

Coomassie brilliant blue

FTIR:

Fourier transform infra red

pHMB:

para-Hydroxymercuricbenzoate

HPLC:

High performance liquid chromatography

SDS–PAGE:

Sodium dodecyl sulphate polyacrylamide gel electrophoresis

UV–VIS:

Ultraviolet visible

GHG-1:

Goat heart galectin-1

PBS:

Phosphate buffered saline

CRD:

Carbohydrate recognition domain

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Acknowledgments

The financial assistance in the form of scholarships to the first author (GMA) 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 facilities. Thanks are due to all friends and fellows for their critical suggestions during experimental set up and manuscript preparation.

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Authors and Affiliations

  1. Amity Institute of Biotechnology (AIB), Amity University Uttar Pradesh (AUUP), Lucknow, 206010, Uttar Pradesh, India

    Ghulam Md Ashraf, Ausaf Ahmad & Rajnish Kumar

  2. Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, 202002, Uttar Pradesh, India

    Ghulam Md Ashraf, Naheed Banu & Ausaf Ahmad

  3. Department of Botany, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, 202002, Uttar Pradesh, India

    Lamabam Peter Singh

Authors
  1. Ghulam Md Ashraf
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  2. Naheed Banu
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  3. Ausaf Ahmad
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  4. Lamabam Peter Singh
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  5. Rajnish Kumar
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Correspondence to Ghulam Md Ashraf.

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Ashraf, G.M., Banu, N., Ahmad, A. et al. Purification, Characterization, Sequencing and Biological Chemistry of Galectin-1 Purified from Capra hircus (goat) Heart. Protein J 30, 39–51 (2011). https://doi.org/10.1007/s10930-010-9300-2

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  • DOI: https://doi.org/10.1007/s10930-010-9300-2

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