The Protein Journal

, 28:391 | Cite as

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

  • Sagarika Biswas
  • Praveen Agrawal
  • Ashish Saroha
  • Hasi R. Das
Article
First Online:

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).

Keywords

Sesbania aculeata Stem lectin 2D gel electrophoresis Isoforms Peptide sequencing MALDI-TOF HPAEC-PAD 

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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Notes

Acknowledgments

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

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Sagarika Biswas
    • 1
  • Praveen Agrawal
    • 1
  • Ashish Saroha
    • 1
  • Hasi R. Das
    • 1
  1. 1.Division of Proteomics and Structural Biology, Institute of Genomics & Integrative BiologyDelhi University CampusDelhiIndia

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