title: Affinity chromatography: a valuable strategy to isolate substrates of methionine sulfoxide reductases?
abstract: Reactive oxygen species fulfill key roles in development and signaling, but lead at high concentration to damage in macromolecules. In proteins, methionine (Met) is particularly prone to oxidative modification and can be oxidized into Met sulfoxide (MetO). MetO reduction is catalyzed by specialized enzymes, termed methionine sulfoxide reductases (MSRs), involved in senescence and protection against diseases and environmental constraints. The precise physiological functions of MSRs remain often elusive because of very poor knowledge of their substrates. In this study, affinity chromatography was used to isolate partners of Arabidopsis thaliana plastidial methionine sulfoxide reductase B1 (MSRB1). Twenty-four proteins involved in photosynthesis, translation, and protection against oxidative stress, as well as in metabolism of sugars and amino acids, were identified. Statistical analysis shows that the abundance of MSRB1 partners in chromatography affinity samples is proportional to Met content. All proteins, for which structural modeling was feasible, display surface-exposed Met and are thus potentially susceptible to oxidation. Biochemical analyses demonstrated that H(2)O(2) treatment actually converts several MSRB1-interacting proteins into MSRB substrates. In consequence, we propose that affinity chromatography constitutes an efficient tool to isolate physiological targets of MSRs.
authors: Lab Ecophysiol Molecul Plantes, Saint-Paul-lez-Durance, France.
authors: Lionel
authors: L
authors: Tarrago
authors: Sylvie
authors: S
authors: Kieffer-Jaquinod
authors: Tiphaine
authors: T
authors: Lamant
authors: Marl Ne
authors: MN
authors: Marcellin
authors: J R Me
authors: JR
authors: Garin
authors: Nicolas
authors: N
authors: Rouhier
authors: Pascal
authors: P
authors: Rey
mesh_terms: D017360:Arabidopsis / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D060365:Chloroplast Proteins / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D051150:Methionine Sulfoxide Reductases / Q000378:metabolism*; D010084:Oxidation-Reduction; D010444:Peptide Elongation Factor Tu / Q000378:metabolism; D011485:Protein Binding
chemical_list: D029681:Arabidopsis Proteins; D060365:Chloroplast Proteins; D051150:Methionine Sulfoxide Reductases; D010444:Peptide Elongation Factor Tu