title: A flowchart to analyze protease activity in plant mitochondria.
abstract: Proteases are one of the most abundant classes of enzymes and are involved in a plethora of biological processes in many cellular compartments, including the mitochondria. To understand the role of proteases is essential to determine their substrate repertoire, preferably in an in vivo setting. In this chapter we describe general guidelines to analyze protease activity using several strategies, from in-gel analysis to mass spectrometry mapping of the cleavage site(s) and fluorogenic probes that can easily be used in vivo. To exemplify this flowchart, we used the recently characterized organellar oligopeptidase of Arabidopsis (Arabidopsis thaliana), an enzyme that takes part in degradation of short peptides within mitochondria and chloroplasts.
authors.affiliation: Department of Biochemistry and Biophysics, Arrhenius Laboratories for Natural Sciences, Stockholm University, Svante Arrhenius väg 16C, Stockholm, 106 91, Sweden, pedro.teixeira@dbb.su.se.
authors.forename: Pedro F
authors.initials: PF
authors.lastname: Teixeira
authors.forename: Rui M
authors.initials: RM
authors.lastname: Branca
authors.forename: Beata
authors.initials: B
authors.lastname: Kmiec
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Glaser
mesh_terms: D017360:Arabidopsis / Q000737:chemistry / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000032:analysis / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D057075:Enzyme Assays / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008928:Mitochondria / Q000737:chemistry / Q000201:enzymology*; D008958:Models, Molecular; D010447:Peptide Hydrolases / Q000032:analysis / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D029681:Arabidopsis Proteins; D010447:Peptide Hydrolases