Stable isotope labeling by amino acids in cell culture (SILAC) combined with high-resolution mass spectrometry is a quantitative strategy for the comparative analysis of (sub)proteomes. It is based on the metabolic incorporation of stable isotope-coded amino acids during growth of cells or organisms. Here, complete labeling of proteins with the amino acid(s) selected for incorporation needs to be guaranteed to enable accurate quantification on a proteomic scale. Wild-type strains of baker's yeast (Saccharomyces cerevisiae ), which is a widely accepted and well-studied eukaryotic model organism, are generally able to synthesize all amino acids on their own (i.e., prototrophic). To render them amenable to SILAC, auxotrophies are introduced by genetic manipulations. We addressed this limitation by developing a generic strategy for complete "native" labeling of prototrophic S. cerevisiae with isotope-coded arginine and lysine, referred to as "2nSILAC". It allows for directly using and screening several genome-wide yeast mutant collections that are easily accessible to the scientific community for functional proteomic studies but are based on prototrophic variants of S. cerevisiae.
%0 Book Section
%1 dannenmaier2nSILACQuantitativeProteomics2021a
%A Dannenmaier, Stefan
%A Oeljeklaus, Silke
%A Warscheid, Bettina
%C United States
%D 2021
%J Methods in molecular biology (Clifton, N.J.)
%K transport
%P 253--270
%R 10.1007/978-1-0716-1024-4_18
%T 2nSILAC for Quantitative Proteomics of Prototrophic Baker's Yeast.
%V 2228
%X Stable isotope labeling by amino acids in cell culture (SILAC) combined with high-resolution mass spectrometry is a quantitative strategy for the comparative analysis of (sub)proteomes. It is based on the metabolic incorporation of stable isotope-coded amino acids during growth of cells or organisms. Here, complete labeling of proteins with the amino acid(s) selected for incorporation needs to be guaranteed to enable accurate quantification on a proteomic scale. Wild-type strains of baker's yeast (Saccharomyces cerevisiae ), which is a widely accepted and well-studied eukaryotic model organism, are generally able to synthesize all amino acids on their own (i.e., prototrophic). To render them amenable to SILAC, auxotrophies are introduced by genetic manipulations. We addressed this limitation by developing a generic strategy for complete "native" labeling of prototrophic S. cerevisiae with isotope-coded arginine and lysine, referred to as "2nSILAC". It allows for directly using and screening several genome-wide yeast mutant collections that are easily accessible to the scientific community for functional proteomic studies but are based on prototrophic variants of S. cerevisiae.
@inbook{dannenmaier2nSILACQuantitativeProteomics2021a,
abstract = {Stable isotope labeling by amino acids in cell culture (SILAC) combined with high-resolution mass spectrometry is a quantitative strategy for the comparative analysis of (sub)proteomes. It is based on the metabolic incorporation of stable isotope-coded amino acids during growth of cells or organisms. Here, complete labeling of proteins with the amino acid(s) selected for incorporation needs to be guaranteed to enable accurate quantification on a proteomic scale. Wild-type strains of baker's yeast (Saccharomyces cerevisiae ), which is a widely accepted and well-studied eukaryotic model organism, are generally able to synthesize all amino acids on their own (i.e., prototrophic). To render them amenable to SILAC, auxotrophies are introduced by genetic manipulations. We addressed this limitation by developing a generic strategy for complete "native" labeling of prototrophic S. cerevisiae with isotope-coded arginine and lysine, referred to as "2nSILAC". It allows for directly using and screening several genome-wide yeast mutant collections that are easily accessible to the scientific community for functional proteomic studies but are based on prototrophic variants of S. cerevisiae.},
added-at = {2024-05-17T13:01:35.000+0200},
address = {United States},
author = {Dannenmaier, Stefan and Oeljeklaus, Silke and Warscheid, Bettina},
biburl = {https://www.bibsonomy.org/bibtex/2cb187e1f6de1b31a476ba39f885a100d/warscheidlab},
doi = {10.1007/978-1-0716-1024-4_18},
interhash = {49426f9bbd25c9361deeb6e449109a5d},
intrahash = {cb187e1f6de1b31a476ba39f885a100d},
issn = {1940-6029 1064-3745},
journal = {Methods in molecular biology (Clifton, N.J.)},
keywords = {transport},
langid = {english},
pages = {253--270},
pmid = {33950496},
timestamp = {2024-05-17T13:56:18.000+0200},
title = {{{2nSILAC}} for {{Quantitative Proteomics}} of {{Prototrophic Baker}}'s {{Yeast}}.},
volume = 2228,
year = 2021
}