Dataset Information

MEERCAT: Multiplexed Efficient Cell Free Expression of Recombinant QconCATS for Large Scale Absolute Proteome Quantification C4PR_LIV

ABSTRACT: A major challenge in proteomics is the absolute accurate quantification of large numbers of proteins. QconCATs, artificial proteins that are concatenations of multiple standard peptides, are well established as an efficient means to generate standards for proteome quantification. Previously, QconCATs have been expressed in bacteria, but we now describe QconCAT expression in a robust, cell-free system. The new expression approach rescues QconCATs that previously were unable to be expressed in bacteria and can reduce the incidence of proteolytic damage to QconCATs. Moreover, it is possible to co-synthesise QconCATs in a highly-multiplexed translation reaction, co-expressing tens or hundreds of QconCATs simultaneously. By obviating bacterial culture and through the gain of high level multiplexing, it is now possible to generate tens of thousands of standard peptides in a matter of weeks, rendering absolute quantification of a complex proteome highly achievable in a reproducible, broadly deployable system.

INSTRUMENT(S):

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

SUBMITTER: Victoria Harman

LAB HEAD: Robert Jeffrey Beynon

PROVIDER: PXD007949 | Pride | 2017-10-27

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	11_1_11_2_export_for_mascot_try2.mgf	Mgf
	11_3_12_1_export_for_mascot_try2.mgf	Mgf
	Coexpression_11_1_11_2_LIGHT_combined_reps_1and2.mzid.gz	Mzid
	Coexpression_11_1_LIGHT_replicate_1.raw	Raw
	Coexpression_11_2_LIGHT_replicate_2.raw	Raw

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Publications

MEERCAT: Multiplexed Efficient Cell Free Expression of Recombinant QconCATs For Large Scale Absolute Proteome Quantification.

Takemori Nobuaki N Takemori Ayako A Tanaka Yuki Y Endo Yaeta Y Hurst Jane L JL Gómez-Baena Guadalupe G Harman Victoria M VM Beynon Robert J RJ

Molecular & cellular proteomics : MCP 20171020 12

A major challenge in proteomics is the absolute accurate quantification of large numbers of proteins. QconCATs, artificial proteins that are concatenations of multiple standard peptides, are well established as an efficient means to generate standards for proteome quantification. Previously, QconCATs have been expressed in bacteria, but we now describe QconCAT expression in a robust, cell-free system. The new expression approach rescues QconCATs that previously were unable to be expressed in bac ...[more]

PMID: 29055021

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