Plant Proteases and Plant Cell Death

Expression and purification of the type I metacaspase from a cryptophyte Guillardia theta, GtMCA-I.- Expression and purification of the type II metacaspase from a unicellular green alga Chlamydomonas reinhardtii.- Expression in Escherichia coli, refolding, and purification of plant aspartic proteases.- Production of functional plant legumain proteases using the Leishmania tarentolae expression system.- Purification of His-tagged proteases from the apoplast of agroinfiltrated N. benthamiana.- Identification of cognate protease/substrate pairs by use of class-specific inhibitors.- Peptide backbone modifications for the assessment of cleavage site relevance in precursors of signaling peptides.- Detection of apoplastic protease inhibitors using convolution activity based proteome profiling.- Cysteine reactivity profiling to unveil redox regulation in phytopathogens.- Determination of caspase-like activities in roots by the use of fluorogenic substrates.- Detection of damage-activated metacaspase activity by Western blot in plants.- Sensitive plant N-terminome profiling with HUNTER.- Profiling sequence specificity of proteolytic activities using proteome-derived peptide libraries.- Monitoring Cell Death via Ion leakage and PAM Fluorometry.- Measuring and perturbing ferroptosis in plants.- Detection and quantification of the hypersensitive response cell death in Arabidopsis thaliana.- Monitoring of ATG4 protease activity during autophagy in the model microalga Chlamydomonas reinhardtii.- Oxidative stress in roots – detection of lipid peroxidation and total antioxidative capacity.- Imaging of potassium and calcium distribution in plant tissues and cells to monitor stress response and programmed cell death.- Monitoring Pseudomonas syringae growth in agroinfiltrated leaves: the ‘agromonas’ assay.- Analysis of virus spread around the cell death zone at spatiotemporal resolutionusing confocal microscopy.- A user guide to validation, annotation and evaluation of N-terminome datasets with MANTI.- Exploring posttranslational modifications with the Plant PTM Viewer.