title: Preparation of Arabidopsis thaliana seedling proteomes for identifying metacaspase substrates by N-terminal COFRADIC.
abstract: Proteome-wide discovery of in vivo metacaspase substrates can be obtained by positional proteomics approaches such as N-terminal COFRADIC, for example by comparing the N-terminal proteomes (or N-terminomes) of wild-type plants to transgenic plants not expressing a given metacaspase. In this chapter we describe a protocol for the preparation of plant tissue proteomes, including differential isotopic labelling allowing for a comparison of in vivo N-terminomes that serves as the starting point for N-terminal COFRADIC studies.
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Liana
authors.initials: L
authors.lastname: Tsiatsiani
authors.forename: Simon
authors.initials: S
authors.lastname: Stael
authors.forename: Petra
authors.initials: P
authors.lastname: Van Damme
authors.forename: Frank
authors.initials: F
authors.lastname: Van Breusegem
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
mesh_terms: D017360:Arabidopsis / Q000201:enzymology*; D020169:Caspases / Q000096:biosynthesis* / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D013058:Mass Spectrometry; D008967:Molecular Biology / Q000379:methods*; D020543:Proteome; D036226:Seedlings / Q000201:enzymology
chemical_list: D020543:Proteome; D020169:Caspases