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Identification of protein complexes with quantitative proteomics in S. cerevisiae
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SILAC yeast: from labeling to comprehensive proteome quantification.

Lyris M F de Godoy1

  • 1Instituto Carlos Chagas, Fiocruz Parana, Rua Prof. Algacyr Munhoz Mader 3775, 81350-010, Curitiba, PR, Brazil, lmfgodoy@fiocruz.br.

Methods in Molecular Biology (Clifton, N.J.)
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Summary

This chapter details Stable Isotope Labeling with Amino acids in Cell culture (SILAC) for yeast proteomics. It provides quality checks and workflows for comprehensive yeast proteome quantification using mass spectrometry.

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Area of Science:

  • Proteomics
  • Biochemistry
  • Yeast biology

Background:

  • Quantitative proteomics enables large-scale analysis of cellular states.
  • Stable Isotope Labeling with Amino acids in Cell culture (SILAC) is a powerful technique for protein quantification.
  • SILAC has broad applications across diverse biological systems.

Purpose of the Study:

  • To provide detailed instructions for SILAC labeling in yeast.
  • To outline quality control measures for labeling efficiency.
  • To present comprehensive workflows for yeast proteome quantification.

Main Methods:

  • Stable Isotope Labeling with Amino acids in Cell culture (SILAC) protocol optimization for yeast.
  • Quality control assays for assessing labeling efficiency.
  • Mass spectrometry-based workflows for quantitative proteomics.

Main Results:

  • Established detailed SILAC labeling procedures for yeast.
  • Developed quality checks to ensure high labeling efficiency.
  • Presented two complete workflows for near-complete yeast proteome quantification.

Conclusions:

  • SILAC is a robust method for yeast proteome analysis.
  • The provided protocols and workflows facilitate broad research applications.
  • This chapter serves as a valuable resource for yeast researchers.