Exploitation of Endogenous Protease Activity in Raw Mastitic Milk by MALDI-TOF/TOF
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- 作者：Anna Napoli ; Donatella Aiello ; Leonardo Di Donna ; Henrrik Prendushi ; Giovanni Sindona
- 刊名：Analytical Chemistry
- 出版年：2007
- 出版时间：August 1, 2007
- 年：2007
- 卷：79
- 期：15
- 页码：5941 - 5948
- 全文大小：566K
- 年卷期：v.79,no.15(August 1, 2007)
- ISSN：1520-6882
文摘
The proteic profiling of bovine milk produced by cows withsubclinical mastitis was obtained by MALDI mass spectrometry. A simple procedure of chemical fractionationof raw milk was developed, whereby less complex fractions of proteins were obtained prior to mass spectrometric and SDS-PAGE analysis. The profiles of milk proteinsthus obtained could allow the identification of either earlymarkers of the acute phase of mastitis or endogenouspeptide of innate immune response. The activity of theendogenous proteases in raw milk produced from eachquarter of healthy and mastic cows was therefore assayedover 24-, 48-, 96-, and 216-h incubation at 37 
C at bothphysiological and acid pH. Sequence-specific peptideswere identified for each fraction by MS/MS experiments,and all tandem mass spectra were evaluated using MASCOT database searching. The results show a specificproteolytic activity of endogenous enzyme toward 
-caseinprecursor (P02666), 
-S2-casein (P02663), -S1-casein(P02662), and 
-casein (P02668).
C at bothphysiological and acid pH. Sequence-specific peptideswere identified for each fraction by MS/MS experiments,and all tandem mass spectra were evaluated using MASCOT database searching. The results show a specificproteolytic activity of endogenous enzyme toward -caseinprecursor (P02666), -S2-casein (P02663), -S1-casein(P02662), and -casein (P02668).