Surface-Based and Mass Spectrometric Approaches to Deciphering Sugar–Protein Interactions in a Galactose-Specific Agglutinin

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2012
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Jiménez-Castells, Carmen
Defaus, Sira
Andreu, David
Gutiérrez-Gallego, Ricardo
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Analytical Chemistry. 2012, 84(15), pp. 6515-6520. ISSN 0003-2700. eISSN 1520-6882. Available under: doi: 10.1021/ac300766z
Zusammenfassung

Interest in powerful, nanosized tools to analyze in detail glycan–protein interactions has increased significantly over recent years. Here, we report two complementary approaches to characterize such interactions with high sensitivity, low sample consumption, and without the need for sample labeling, namely, surface plasmon resonance (SPR) and an approach that combines limited proteolysis and mass spectrometry. Combination of these two approaches to investigate glycan–protein interactions allows (1) to characterize interactions through kinetic and thermodynamic parameters, (2) to capture efficiently the carbohydrate-binding protein, and (3) to identify the interacted protein and its carbohydrate binding site by mass spectrometry. As a proof of principle, the interaction of the galactose-specific legume lectin Erythrina cristagalli agglutinin with several sugars has been characterized in-depth by means of these two approaches.

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ISO 690JIMÉNEZ-CASTELLS, Carmen, Sira DEFAUS, Adrian MOISE, Michael PRZYBYLSKI, David ANDREU, Ricardo GUTIÉRREZ-GALLEGO, 2012. Surface-Based and Mass Spectrometric Approaches to Deciphering Sugar–Protein Interactions in a Galactose-Specific Agglutinin. In: Analytical Chemistry. 2012, 84(15), pp. 6515-6520. ISSN 0003-2700. eISSN 1520-6882. Available under: doi: 10.1021/ac300766z
BibTex
@article{JimenezCastells2012-08-07Surfa-22619,
  year={2012},
  doi={10.1021/ac300766z},
  title={Surface-Based and Mass Spectrometric Approaches to Deciphering Sugar–Protein Interactions in a Galactose-Specific Agglutinin},
  number={15},
  volume={84},
  issn={0003-2700},
  journal={Analytical Chemistry},
  pages={6515--6520},
  author={Jiménez-Castells, Carmen and Defaus, Sira and Moise, Adrian and Przybylski, Michael and Andreu, David and Gutiérrez-Gallego, Ricardo}
}
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