Detection of protein oxidation products by fluorescence spectroscopy and trilinear data decomposition: Proof of concept
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Detection of protein oxidation products by fluorescence spectroscopy and trilinear data decomposition : Proof of concept. / Bevilacqua, Marta; Engholm-Keller, Kasper; Risum, Anne Bech; Rinnan, Åsmund; Lund, Marianne N.
In: Food Chemistry, Vol. 396, 133732, 2022.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Detection of protein oxidation products by fluorescence spectroscopy and trilinear data decomposition
T2 - Proof of concept
AU - Bevilacqua, Marta
AU - Engholm-Keller, Kasper
AU - Risum, Anne Bech
AU - Rinnan, Åsmund
AU - Lund, Marianne N.
N1 - Publisher Copyright: © 2022 The Author(s)
PY - 2022
Y1 - 2022
N2 - Current analytical methods studying protein oxidation modifications require laborious sample preparation and chromatographic methods. Fluorescence spectroscopy is an alternative, as many protein oxidation products are fluorescent. However, the complexity of the signal causes misinterpretation and quantification errors if single emission spectra are used. Here, we analyzed the entire fluorescence excitation-emission matrix using the trilinear decomposition method parallel factor analysis (PARAFAC). Two sample sets were used: a calibration set based on known mixtures of tryptophan, tyrosine, and four oxidation products, and a second sample set of oxidized protein solutions containing UV-illuminated β-lactoglobulin. The PARAFAC model succeeded in resolving the signals of the model systems into the pure fluorophore components and estimating their concentrations. The estimated concentrations for the illuminated β-lactoglobulin samples were validated by liquid chromatography-mass spectrometry. Our approach is a promising tool for reliable identification and quantification of fluorescent protein oxidation products, even in a complex protein system.
AB - Current analytical methods studying protein oxidation modifications require laborious sample preparation and chromatographic methods. Fluorescence spectroscopy is an alternative, as many protein oxidation products are fluorescent. However, the complexity of the signal causes misinterpretation and quantification errors if single emission spectra are used. Here, we analyzed the entire fluorescence excitation-emission matrix using the trilinear decomposition method parallel factor analysis (PARAFAC). Two sample sets were used: a calibration set based on known mixtures of tryptophan, tyrosine, and four oxidation products, and a second sample set of oxidized protein solutions containing UV-illuminated β-lactoglobulin. The PARAFAC model succeeded in resolving the signals of the model systems into the pure fluorophore components and estimating their concentrations. The estimated concentrations for the illuminated β-lactoglobulin samples were validated by liquid chromatography-mass spectrometry. Our approach is a promising tool for reliable identification and quantification of fluorescent protein oxidation products, even in a complex protein system.
KW - 3,4-dihydroxy-L-phenylalanine (PubChem CID: 6047)
KW - 5-hydroxy-L-Trp (PubChem CID: 439280)
KW - dityrosine (PubChem CID: 107904)
KW - Excitation-Emission matrix
KW - Fluorescence spectroscopy
KW - N-formylkynurenine (PubChem CID: 910)
KW - Parallel Factor Analysis (PARAFAC)
KW - Protein oxidation
KW - Second order advantage
KW - Tryptophan (PubChem CID: 6305)
KW - tyrosine (PubChem CID: 1153)
KW - β-lactoglobulin
U2 - 10.1016/j.foodchem.2022.133732
DO - 10.1016/j.foodchem.2022.133732
M3 - Journal article
C2 - 35872499
AN - SCOPUS:85134584959
VL - 396
JO - Food Chemistry
JF - Food Chemistry
SN - 0308-8146
M1 - 133732
ER -
ID: 316395888