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For database search the following parameters were used: two missed cleavages, carbamidomethylation of cysteine (as fixed modification) and oxidation of methionine (as variable modification).
Carbamidomethylation of cysteine was set as fixed modification and oxidation of methionines as variable modification.
Carbamidomethylcysteine was selected as a fixed modification and oxidation of methionine as a variable modification.
Carbamidomethylation (C) was set as a fixed modification, and oxidation (M), phosphorylation (S, T and Y) and deamidation (N) were set as variable modifications.
The following parameters were used for the database search: MS and MS/MS accuracies were set to <0.6Da, trypsin/P as an enzyme, missed cleavages 1, carbamidomethylation of cysteine as fixed modification, and oxidation of methionine as a variable modification.
CID spectra were interpreted by Phenyx software (Genebio SA, Geneva, Switzerland) using carboamidomethylation of cysteine as a fixed modification, and oxidation of methionine and EPG on glutamic acid as variable modification.
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The MASCOT search parameters were as follows: (i) taxonomy: Arabidopsis; (ii) potential modifications: carbamidomethyl (C) and GlyGly (K) as fixed modifications, and oxidation (M) as a variable modification; (iii) max missed cleavage: 1; (iv) peptide MS tolerance: ±10 ppm; and (v) fragment mass tolerance: ± 0.5 Da.
Other parameters include methylthiol modification of cysteine, iTRAQ labels at the peptide N-terminus and Lysine residues as static modifications and oxidation of methionine as variable modification.
Carbamidomethylation of Cys (C) residues were set as fixed modifications, and oxidation of Met and acetylation of N-terminal G1 and K2 residues were set as variable modifications.
For the global proteomic dataset, carbamidomethylation of cysteine and iTRAQ 4-plex modification of peptide N-termini were set as fixed modifications, and oxidation of methionine and iTRAQ 4-plex modification of Lys were set as variable modifications.
Further settings used were the following: trypsin with 1 missed cleavage; carbamidomethylation on cysteine and iTRAQ-8plex on lysine and N-terminal as fixed modifications; and oxidation of methionine as variable modification.
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