Dataset Information

Absolute Quantification of Yeast Proteins Using Synthetic Peptide Standards and TMT labeling

ABSTRACT: The absolute protein amounts for the selected yeast ribosomal proteins (RPs) were determined using the standard peptides and TMT labelling. The range of the standard tryptic peptides were selected and synthesized based on the preliminary data-dependent experiments. The synthetic peptides were mixed and four aliquots containing different amounts of the peptides were labelled each with a different TMT labelling reagent. The yeast cell lysate was digested and labelled in parallel. The labelled cellular and standard samples were combined, and the absolute peptide amounts in the lysate were calculated by comparison to the known amounts in the standard TMT channels.

INSTRUMENT(S): Orbitrap Fusion

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

SUBMITTER: Egor Vorontsov

LAB HEAD: Jens Nielsen

PROVIDER: PXD015025 | Pride | 2020-03-13

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	3118_RefR2824_Amounts_ManuallyCheckedPSMs.xlsx	Xlsx
	3118_Stand_Trypsin.fasta	Fasta
	Fusion_190408_101.raw	Raw
	Fusion_190408_80.raw	Raw
	Fusion_190408_95.raw	Raw

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Publications

Nitrogen limitation reveals large reserves in metabolic and translational capacities of yeast.

Yu Rosemary R Campbell Kate K Pereira Rui R Björkeroth Johan J Qi Qi Q Vorontsov Egor E Sihlbom Carina C Nielsen Jens J

Nature communications 20200420 1

Cells maintain reserves in their metabolic and translational capacities as a strategy to quickly respond to changing environments. Here we quantify these reserves by stepwise reducing nitrogen availability in yeast steady-state chemostat cultures, imposing severe restrictions on total cellular protein and transcript content. Combining multi-omics analysis with metabolic modeling, we find that seven metabolic superpathways maintain >50% metabolic capacity in reserve, with glucose metabolism maint ...[more]

PMID: 32312967

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