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Influence of various endogenous and artefact modifications on large‐scale proteomics analysis
Author(s) -
Bienvenut Willy V.,
Sumpton David,
Lilla Sergio,
Martinez Aude,
Meinnel Thierry,
Giglione Carmela
Publication year - 2012
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.6474
Subject(s) - chemistry , peptide , acetylation , proteome , proteomics , posttranslational modification , computational biology , phosphorylation , identification (biology) , biochemistry , combinatorial chemistry , chromatography , enzyme , biology , botany , gene
RATIONALE Some large‐scale proteomics studies in which strong cation exchange chromatography has been applied are used to determine proteomes and post‐translational modification dynamics. Although such datasets favour the characterisation of thousands of modified peptides, e.g., phosphorylated and N‐α‐acetylated, a large fraction of the acquired spectra remain unexplained by standard proteomics approaches. Thus, advanced data processing allows characterisation of a significant part of these unassigned spectra. METHODS Our recent investigation of the N‐α‐acetylation status of plant proteins gave a dataset of choice to investigate further the in‐depth characterisation of peptide modifications using Mascot tools associated with relevant validation processes. Such an approach allows to target frequently occurring modifications such as methionine oxidation, phosphorylation or N‐α‐acetylation, but also the less usual peptide cationisation. Finally, this dataset offers the unique opportunity to determine the overall influence of some of these modifications on the identification score. RESULTS Although methionine oxidation has no influence and tends to favour the characterisation of protein N‐terminal peptides, peptide alkalinisation shows an adverse effect on peptide average score. Nevertheless, peptide cationisation appears to favour the characterisation of protein C‐terminal peptides with a limited to no direct influence on the identification score. Unexpectedly, our investigation reveals the unfortunate combination of the molecular weight of N‐α‐acetylation and potassium cation that mimics the mass increment of a phosphorylation group. CONCLUSIONS Since these characterisations rely upon computational treatment associated with statistical validation approaches such as 'False discovery rates' calculation or post‐translational modification position validation, our investigation highlights the limitation of such treatment which is biased by the initial searched hypotheses. Copyright © 2012 John Wiley & Sons, Ltd.