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Absolute quantification of budding yeast proteome with PCS method

ABSTRACT: Absolute quantification of proteome is one of the most important tasks in proteomic research. The aim in this analysis is providing proof-of-principle of our originally developed strategy for absolute quantification of multiple proteins present at various concentrations in a cell, using stable isotope-labeled peptide concatenated standard (PCS). This study also provides insights into structures of regulation of metabolic pathway and into biological mechanisms for regulation of expressed protein abundance.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Culture

SUBMITTER: Keiji Kito

LAB HEAD: Keiji Kito

PROVIDER: PXD003084 | Pride | 2016-10-06

REPOSITORIES: Pride

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A strategy for absolute proteome quantification with mass spectrometry by hierarchical use of peptide-concatenated standards.

Kito Keiji K Okada Mitsuhiro M Ishibashi Yuko Y Okada Satoshi S Ito Takashi T

Proteomics 20160428 10

The accurate and precise absolute abundance of proteins can be determined using mass spectrometry by spiking the sample with stable isotope-labeled standards. In this study, we developed a strategy of hierarchical use of peptide-concatenated standards (PCSs) to quantify more proteins over a wider dynamic range. Multiple primary PCSs were used for quantification of many target proteins. Unique "ID-tag peptides" were introduced into individual primary PCSs, allowing us to monitor the exact amounts ...[more]

PMID: 27030420

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