Determination of exposed sulphydryl groups in heated β-lactoglobulin A using IAEDANS and mass spectrometry
dc.contributor.author | Kehoe, Joseph James | |
dc.contributor.author | Brodkorb, Andre | |
dc.contributor.author | Molle, Daniel | |
dc.contributor.author | Yokoyama, Emilie | |
dc.contributor.author | Famelart, Marie-Helene | |
dc.contributor.author | Bouhallab, Said | |
dc.contributor.author | Morris, Edwin R | |
dc.contributor.author | Croguennec, Thomas | |
dc.date.accessioned | 2014-01-21T14:59:03Z | |
dc.date.available | 2014-01-21T14:59:03Z | |
dc.date.issued | 25/07/2007 | |
dc.identifier.citation | Joseph J. Kehoe, André Brodkorb, Daniel Mollé, Emilie Yokoyama, Marie-Héléne Famelart, Saíd Bouhallab, Edwin R. Morris, and Thomas Croguennec. Determination of Exposed Sulfhydryl Groups in Heated β-Lactoglobulin A Using IAEDANS and Mass Spectrometry. J. Agric. Food Chem., 2007, 55 (17), 7107–7113. DOI: 10.1021/jf070397r | en_GB |
dc.identifier.issn | 0021-8561 | |
dc.identifier.uri | http://hdl.handle.net/11019/512 | |
dc.description | peer-reviewed | en_GB |
dc.description.abstract | This paper takes a new approach to determining which sulfhydryl groups are exposed during the heat denaturation of bovine β-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 β-lactoglobulin A it was found that cysteine 66 and cysteine 160 were predominantly exposed regardless of the length of exposure to heat. | en_GB |
dc.description.sponsorship | Travel costs associated with exchanges of scientists between the authors’ laboratories were co-funded by the Enterprise Ireland / French Ministry for Foreign Affairs Ulysses Exchange Research Program. The cost of project work was supported by the Food Institutional Research Measure (FIRM) of the National Development Plan 2000-2006 (Ireland). J. Kehoe was funded by the Teagasc Walsh Fellowship Scheme | |
dc.language.iso | en | en_GB |
dc.publisher | American Chemical Society | en_GB |
dc.relation.ispartofseries | Journal of Agricultural and Food Chemistry;vol 55 | |
dc.subject | β-Lactoglobulin | en_GB |
dc.subject | Free sulfhydryl | en_GB |
dc.subject | IAEDANS | en_GB |
dc.subject | MALDI mass spectrometry | en_GB |
dc.title | Determination of exposed sulphydryl groups in heated β-lactoglobulin A using IAEDANS and mass spectrometry | en_GB |
dc.type | Article | en_GB |
dc.identifier.rmis | NFFM-0203-4764 | |
dc.identifier.rmis | MDDT-0103-5452 | |
dc.identifier.doi | http://dx.doi.org/10.1021/jf070397r | |
dc.contributor.sponsor | Teagasc Walsh Fellowship Programme | |
dc.contributor.sponsor | Enterprise Ireland | |
dc.contributor.sponsor | Department of Agriculture Food and the Marine | |
dc.contributor.sponsor | French Ministry for Foreign Affairs | |
refterms.dateFOA | 2018-01-12T07:55:10Z |