Award details

Extending absolute quantification to the proteome-wide scale.

Reference	BB/C007735/1
Principal Investigator / Supervisor	Professor Simon James Gaskell
Co-Investigators / Co-Supervisors
Institution	The University of Manchester
Department	Chemistry
Funding type	Research
Value (£)	226,724
Status	Completed
Type	Research Grant
Start date	01/04/2005
End date	28/02/2009
Duration	47 months

Abstract

One of the unsolved challenges in proteomics is that of absolute, rather than relative quantification. We propose a novel solution to this challenge, by creating artificial genes consisting of a concatenated series of peptides, such that each peptide acts as a signature quantification Q-peptide for one protein. A typical concatamer, designed from an optimal assembly of Q-peptides, would be a standard for approx 50 proteins. The concatamer also contains purification and quantification motifs at the N and C termini, so that it is possible to purify the bacterially expressed protein and then quantify the intact protein accurately. After proteolysis, the released Q-peptides than act as standards for the cognate proteins. The Q-peptides can be used directly, stable isotope labelled and added directly to the analyte, or indirectly, in which they are used to deliver an accurate, and global quantification of the proteome in a type strain (itself stable isotope labelled) and which can then be used in all future studies of that organism. In this programme, we will develop optimal strategies for concatamer design, expression and manipulation, and establish the best modalities for their use. We will address the issues of dynamic range, demonstrate the applicability of this approach to selective subproteomes and develop advanced MS strategies to enhance selectivity and sensitivity of the MS quantification step. Joint with BB/C007433/1

Summary

unavailable

Committee	Closed Committee - Engineering & Biological Systems (EBS)
Research Topics	Technology and Methods Development
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

Associated awards:

BB/C007433/1 Extending absolute quantification to the proteome-wide scale.

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