Different carbon sources affect lifespan and protein redox state during Saccharomyces cerevisiae chronological ageing

Cell Mol Life Sci. 2009 Mar;66(5):933-47. doi: 10.1007/s00018-009-8574-z.

Abstract

In this study, a proteomic approach that combines selective labelling of proteins containing reduced cysteine residues with two-dimensional electrophoresis/mass spectrometry was used to evaluate the redox state of protein cysteines during chronological ageing in Saccharomyces cerevisiae. The procedure was developed on the grounds that biotin-conjugated iodoacetamide (BIAM) specifically reacts with reduced cysteine residues. BIAM-labelled proteins can then be selectively isolated by streptavidin affinity capture. We compared cells grown on 2% glucose in the exponential phase and during chronological ageing and we found that many proteins undergo cysteine oxidation. The target proteins include enzymes involved in glucose metabolism. Both caloric restriction and growth on glycerol resulted in a decrease in the oxidative modification. Furthermore, in these conditions a reduced production of ROS and a more negative glutathione half cell redox potential were observed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Caloric Restriction
  • Carbon / metabolism*
  • Cysteine / metabolism
  • Cytochromes / metabolism
  • Glucose / metabolism
  • Glutathione / metabolism
  • Glycerol / metabolism
  • Mitochondria / metabolism
  • Mitochondria / ultrastructure
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Oxygen Consumption
  • Proteomics / methods
  • Reactive Oxygen Species / metabolism
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / physiology*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / isolation & purification
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Time Factors

Substances

  • Cytochromes
  • Reactive Oxygen Species
  • Saccharomyces cerevisiae Proteins
  • Carbon
  • Glutathione
  • Glucose
  • Cysteine
  • Glycerol