Determination of exposed sulfhydryl groups in heated β-lactoglobulin A using IAEDANS and mass spectrometry - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Journal of Agricultural and Food Chemistry Année : 2007

Determination of exposed sulfhydryl groups in heated β-lactoglobulin A using IAEDANS and mass spectrometry

Résumé

This paper takes a new approach to determining which sulfhydryl groups are exposed during the heat denaturation of bovine â-lactoglobulin A. The sulfhydryl groups exposed after heating were blocked with 5-((((2-iodoacetyl)amino)ethyl)amino)naphthalene-1-sulfonic acid (IAEDANS). The results show that IAEDANS is a suitable blocking agent, and its absorbance at 336 nm enabled the quantification of exposed sulfhydryl groups in a mixture of protein species by gel permeation chromatography. Combined with the specific fragmentation of bound IAEDANS by matrix-assisted laser desorption ionization (MALDI) MS/MS in negative ionization mode, this facilitated the identification of peptides that contained blocked cysteines after enzymatic digestion of the protein. During MALDI MS/MS of the peptides, in positive ionization mode, the IAEDANS molecule remained bound to the cysteines, making it possible to identify exactly which cysteine had been exposed after heating. In â-lactoglobulin A it was found that cysteine 66 and cysteine 160 were predominantly exposed regardless of the length of exposure to heat.

Dates et versions

hal-01454031 , version 1 (02-02-2017)

Identifiants

Citer

Joseph Kehoe, André Brodkorb, Daniel Mollé, Emilie Yokoyama, Marie-Hélène Famelart, et al.. Determination of exposed sulfhydryl groups in heated β-lactoglobulin A using IAEDANS and mass spectrometry. Journal of Agricultural and Food Chemistry, 2007, 55 (17), pp.7107-7113. ⟨10.1021/jf070397r⟩. ⟨hal-01454031⟩
71 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More