skip to main content
Show Results with: Show Results with:

An improved workflow for quantitative N‐terminal charge‐based fractional diagonal chromatography (ChaFRADIC) to study proteolytic events in Arabidopsis thaliana

Venne, A. Saskia et al.

Proteomics. Volume 15:Issue 14 (2015, July); pp 2458-2469 -- Wiley-VCH Verlag

Online access

  • Title:
    An improved workflow for quantitative N‐terminal charge‐based fractional diagonal chromatography (ChaFRADIC) to study proteolytic events in Arabidopsis thaliana
  • Author: Venne, A. Saskia;
    Solari, Fiorella A.;
    Faden, Frederik;
    Paretti, Tomasso;
    Dissmeyer, Nico;
    Zahedi, René P.;
    Van Damme, Petra;
    Gevaert, Kris
  • Found In: Proteomics. Volume 15:Issue 14 (2015, July); pp 2458-2469
  • Journal Title: Proteomics
  • Subjects: Bioinformatics--Periodicals; Genomes--Periodicals; Molecular genetics--Periodicals; Proteins--Separation--Periodicals; Proteomics--Periodicals; Dewey: 572.605
  • Rights: legaldeposit
  • Publication Details: Wiley-VCH Verlag
  • Abstract: <x xml:space="preserve">Abstract</x>

    We applied an extended charge‐based fractional diagonal chromatography (ChaFRADIC) workflow to analyze the N‐terminal proteome of Arabidopsis thaliana seedlings. Using iTRAQ protein labeling and a multi‐enzyme digestion approach including trypsin, GluC, and subtilisin, a total of 200 μg per enzyme, and measuring only one third of each ChaFRADIC‐enriched fraction by LC‐MS, we quantified a total of 2791 unique N‐terminal peptides corresponding to 2249 different unique N‐termini from 1270 Arabidopsis proteins. Our data indicate the power, reproducibility, and sensitivity of the applied strategy that might be applicable to quantify proteolytic events from as little as 20 μg of protein per condition across up to eight different samples. Furthermore, our data clearly reflect the methionine excision dogma as well as the N‐end rule degradation pathway (NERP) discriminating into a stabilizing or destabilizing function of N‐terminal amino acid residues. We found bona fide NERP destabilizing residues underrepresented, and the list of neo N‐termini from wild type samples may represent a helpful resource during the evaluation of NERP substrate candidates. All MS data have been deposited in the ProteomeXchange with identifier PXD001855 (http://proteomecentral.proteomexchange.org/dataset/PXD001855).


  • Identifier: ETOClsidyv978b424c; System Number: LDEAvdc_100025981432.0x000001; Journal ISSN: 1615-9853; 10.1002/pmic.201500014
  • Publication Date: 2015
  • Physical Description: Electronic
  • Shelfmark(s): ELD Digital store

Searching Remote Databases, Please Wait