Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling.

Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling. Research Abstract Details 

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Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling. Abstract Text:

Julia R Smith,Michael Olivier,Andrew S Greene,Julia R Smith,Michael Olivier,Andrew S Greene,

We have developed a method to determine the degree of phosphorylation of a peptide in a complex mixture without enrichment or operation of the mass spectrometer in negative ion mode. Yeast lysate containing known amounts of synthetic peptides (VPQLEIVPNSAEERLHSMK and VPQLEIVPN[pS]AEERLHSMK) was labeled with (16)O and (18)O during hydrolysis. After treatment of one sample with a cocktail of phosphatases, the two samples were pooled. The intensity of the dephosphorylated peptide peaks was used to infer the degree of phosphorylation present before treatment. The linear dynamic range of this method is >10-fold before either of the peptide envelopes becomes indistinguishable from the surrounding noise. Since both the site of posttranslational modification and the proportion of the protein population that is modified are vital in protein function, the employment of this technique will provide a valuable tool for the analysis of the functional implications of protein phosphorylation.

Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling. Publishing Authors By Initials

JR Smith,M Olivier,AS Greene,JR Smith,M Olivier,AS Greene,

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Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Physiological genomics

VOLUME: 31

Page Numbers: 357-63

Journal Abbreviation: Physiol. Genomics

ISSN: 1531-2267

DAY: 7

MONTH: 08

YEAR: 2007

Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling. Information

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LANGUAGE: eng

NlmUniqueID: 9815683

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Grant and Affiliation Information for Relative quantification of peptide phosphorylation in a complex mixture using 18O labeling.

AFFILIATION: Biotechnology and Bioengineering Center, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.

Country: United States

AGENCY: United States NHLBI

GRANT: N01-HV-28182

ACRONYM: HV

MEDLINETA: Physiol Genomics

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