Two-dimensional gel electrophoresis of proteins and peptides in bovine milk

Lindmark-Månsson, Helena; Timgren, Anna; Aldén, Gun; Paulsson, Marie

doi:10.1016/j.idairyj.2004.06.010

Cited by 39 publications

(34 citation statements)

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“…It has been reported that the concentration of denatured whey proteins was only 2.8% in milk pasteurised at 72°C for 26 s, whereas the concentration increased to 8.4% when increasing the temperature to 87°C [27]. Using the more severe heat treatment of 85°C for 30 min resulted in a marked decrease in the concentration and number of proteins in whey as compared to a heat treatment of 65°C for 30 min [18].…”

Section: Proteomic Profiles Of Sweet Whey and Rennet Caseinmentioning

confidence: 94%

Different proteomic profiles of sweet whey and rennet casein obtained after preparation from raw versus heat-treated skimmed milk

Larsen

Wedholm-Pallas

Lindmark-Månsson³

et al. 2010

Dairy Sci. Technol.

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-Proteomic profiles of sweet whey and rennet casein fractions prepared from raw or pasteurised (72°C for 15 s), skimmed milk were studied by proteomics coupled with the detection of protein spots by MALDI TOF mass spectrometry. Proteins were analysed by a modification of the traditional, two-dimensional gel electrophoresis (2-DGE) method under non-reducing running conditions, which potentially permits the visualisation of disulphide-linked protein complexes formed in response to pasteurisation. Separated proteins were stained with Coomassie blue. The relative spot volumes obtained after 2-DGE of fractions from raw or pasteurised milk were compared. A number of different spots in the rennet casein and sweet whey fractions were found to vary in response to pasteurisation. Some of these represented higher molecular mass complexes that increased in the chymosin-precipitated casein fraction, and they were identified by mass spectrometry to contain α S1 -casein. Certain fragments of α S1 -casein, probably generated as a result of chymosin cleavage, increased in the whey after pasteurisation. The whey content of proteose peptone component 3 (PP3) or lactophorin decreased after pasteurisation, which could indicate increased association of PP3 with for example the milk fat globule membrane after pasteurisation. This shows that gel-based proteome analysis can be used in the characterisation of subtle variations in protein composition of milk fractions that occur as a consequence of pasteurisation.proteome analysis / sweet whey / rennet casein / pasteurisation Article published by EDP SciencesRésumé -Différents profils protéomiques obtenus à partir de lactosérum doux et de caséine présure issus de laits écrémés crus ou traités thermiquement. Les profils protéomiques de fractions obtenus à partir de lactosérum doux et de caséine présure de lait écrémé cru ou pasteurisé (72°C, 15 s) ont été étudiés par des analyses protéomiques couplées à une détection des spots protéiques par spectrométrie de masse MALDI TOF. Les protéines ont été analysées par électro-phorèse sur gel bidimensionnelle (EG-2D) avec une modification de la méthode classique, sous conditions non réductrices, permettant la visualisation des complexes de protéines liées par ponts disulphure formés en réponse à la pasteurisation. Les protéines séparées étaient colorées au bleu de Coomassie. Les volumes relatifs des spots obtenus après EG-2D des fractions obtenues à partir de lait cru ou pasteurisé ont été comparés. Un nombre de spots différents était trouvé dans les fractions de caséine présure et de lactosérum doux en réponse à la pasteurisation. Certains spots représen-taient des complexes de masse moléculaire plus élevée qui augmentaient dans la fraction de caséine obtenue par précipitation due à la chymosine et qui contenaient des caséines alpha S1 identifiées par spectrométrie de masse. Certains fragments de caséine alpha S1, probablement obtenus après clivage par la chymosine, augmentaient dans le lactosérum après pasteurisation. La teneur du lactos...

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Section: Proteomic Profiles Of Sweet Whey and Rennet Caseinmentioning

confidence: 94%

Different proteomic profiles of sweet whey and rennet casein obtained after preparation from raw versus heat-treated skimmed milk

Larsen

Wedholm-Pallas

Lindmark-Månsson³

et al. 2010

Dairy Sci. Technol.

Self Cite

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“…As shown in Fig. 2, four protein bands of whey in native-PAGE were re-separated by SDS-PAGE, and were confirmed as BSA, a-LA, b-LGB and b-LGA, respectively, according to their molecular weights and relative contents in whey proteins (Lindmark-Månsson et al, 2005). Additional characterization by SDS-PAGE with excised native-PAGE protein bands would be a helpful tool to identify some thermally unstable enzymes or proteins when the standard protein is unavailable.…”

Section: Application Of Native-pagementioning

confidence: 99%

“…A list of methods has been reported in the literature for the determination of whey protein fractions: (1) electrophoretic techniques using polyacrylamide gels with urea or sodium dodecylsulfate (SDS) and two-dimensional techniques (Lindmark-Månsson, Timgren, Aldén, & Paulsson, 2005;Veloso, Teixeira, Peres, Mendonca, & Ferreira, 2004); (2) high-performance liquid chromatography by ion-exchange, hydrophobic interaction, gel filtration and reversed-phase modes (Bramanti et al, 2004;Lozano, Giraldo, & Romero, 2008;Wedholm et al, 2006); and (3) immunological methods (Chen, Huang, Liu, Li, & Mao, 2004), capillary electrophoreses (Heck et al, 2008;Rehder & McGown, 2001) and electrospray ionization mass spectrometry (Losito et al, 2007). Methods based on immunological and electrophoretic techniques have been proposed to be the most accurate and sensitive techniques (Kaminarides & Koukiassa, 2002).…”

Section: Introductionmentioning

confidence: 99%

Distinction of different heat-treated bovine milks by native-PAGE fingerprinting of their whey proteins

et al. 2010

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“…Two-dimensional gel electrophoresis was performed on whey samples according to Lindmark-Ma˚nsson, Timgren, Alde´n, and Paulsson (2005). Immobilised pH gradient (IPG) strips, Immobiline DryStrip gels (Amersham Biosciences) were used for separation in the first dimension.…”

Section: Two-dimensional Gel Electrophoresismentioning

confidence: 99%

Relationship between somatic cell count, individual leukocyte populations and milk components in bovine udder quarter milk

Lindmark-Månsson¹,

Bränning

Aldén³

et al. 2006

International Dairy Journal

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103

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Two-dimensional gel electrophoresis of proteins and peptides in bovine milk

Cited by 39 publications

References 38 publications

Different proteomic profiles of sweet whey and rennet casein obtained after preparation from raw versus heat-treated skimmed milk

Different proteomic profiles of sweet whey and rennet casein obtained after preparation from raw versus heat-treated skimmed milk

Distinction of different heat-treated bovine milks by native-PAGE fingerprinting of their whey proteins

Relationship between somatic cell count, individual leukocyte populations and milk components in bovine udder quarter milk

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