Different carbon sources affect lifespan and protein redox state during Saccharomyces cerevisiae chronological ageing.
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.
Magherini
F
F
Dipartimento di Scienze Biochimiche Università degli Studi di Firenze, Viale G.Morgagni 50 I-50134, Firenze, Italy. francesca.magherini@unifi.it
Carpentieri
A
A
Amoresano
A
A
Gamberi
T
T
De Filippo
C
C
Rizzetto
L
L
Biagini
M
M
Pucci
P
P
Modesti
A
A
D000595:Amino Acid Sequence; D031204:Caloric Restriction; D002244:Carbon / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D003580:Cytochromes / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism / Q000648:ultrastructure; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D040901:Proteomics / Q000379:methods; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013997:Time Factors
D003580:Cytochromes; D017382:Reactive Oxygen Species; D029701:Saccharomyces cerevisiae Proteins; D002244:Carbon; D005978:Glutathione; D005947:Glucose; D003545:Cysteine; D005990:Glycerol