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Purification and Mass Spectrometric Characterization of Sesbania aculeata (Dhaincha) Stem Lectin

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Abstract

A glucose specific lectin (STA) was isolated from Sesbania aculeata stem by using Sephadex G-50 affinity column chromatography. The lectin is a glycoprotein having 29 kDa subunit molecular weight. Two dimensional gel electrophoresis analysis revealed that the lectin existed in two isomeric forms with varied carbohydrate content as analyzed by high performance anion exchange chromatography-pulsed amperometric detector (HPAEC-PAD). Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and N-terminal sequence (LDSLSFTYNNFE) analysis of this lectin showed 95% homology with stem lectin SL-I (accession no. AJ585523) from peanut plant. The nucleotide sequence of the lectin (STA) was submitted to the gene bank (accession no. EU263636).

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Abbreviations

MALDI-TOF/TOF:

Matrix assisted laser desorption ionization time of flight mass spectrometry

PMSF:

Phenylmethanesulfonyl fluoride

HPAEC-PAD:

High performance anion exchange chromatography-pulsed amperometric detector

HA:

Hemagglutination activity

IEF:

Iso-electric focusing

PNGase F:

Peptide N-glycosidase F

TBS:

Tris buffered saline

ELLBA:

Enzyme linked lectin binding assay

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Acknowledgments

Financial assistance from the Department of Biotechnology, India is gratefully acknowledged.

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Authors and Affiliations

  1. Division of Proteomics and Structural Biology, Institute of Genomics & Integrative Biology, Delhi University Campus, Mall Road, Delhi, 110007, India

    Sagarika Biswas, Praveen Agrawal, Ashish Saroha & Hasi R. Das

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  1. Sagarika Biswas
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  2. Praveen Agrawal
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  3. Ashish Saroha
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  4. Hasi R. Das
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Correspondence to Hasi R. Das.

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Biswas, S., Agrawal, P., Saroha, A. et al. Purification and Mass Spectrometric Characterization of Sesbania aculeata (Dhaincha) Stem Lectin. Protein J 28, 391–399 (2009). https://doi.org/10.1007/s10930-009-9206-z

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  • DOI: https://doi.org/10.1007/s10930-009-9206-z

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