title: Kinetic Analysis of Plant SUMO Conjugation Machinery.
abstract: Plants display a high diversification degree of the SUMO conjugation machinery, which could confer a biological specialization of the different isoforms. For instance, the two essential Arabidopsis SUMO isoforms, SUMO1/2, display the highest conjugation rate when compared to SUMO3 and 5, suggesting that their specific biochemical properties may be linked to their biological specialization. In order to study the biochemical properties of plant SUMO conjugation systems, quantitative biochemical assays must be performed. We will present a detailed protocol for reconstituting an in vitro SUMO conjugation assay covering all steps from protein preparation to assay development.
authors.affiliation: Development Program, CRAG (CSIC-IRTA-UAB-UB), Edifici CRAG-Campus UAB, Bellaterra (Cerdanyola del Vallés), 08193, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Castaño-Miquel
authors.affiliation: Development Program, CRAG (CSIC-IRTA-UAB-UB), Edifici CRAG-Campus UAB, Bellaterra (Cerdanyola del Vallés), 08193, Barcelona, Spain. maria.lois@cragenomica.es.
authors.forename: L Maria
authors.initials: LM
authors.lastname: Lois
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics; D007700:Kinetics; D008967:Molecular Biology / Q000379:methods*; D020033:Protein Isoforms / Q000737:chemistry* / Q000235:genetics; D058207:Sumoylation / Q000235:genetics*
chemical_list: D029681:Arabidopsis Proteins; D020033:Protein Isoforms; C477512:SUM1 protein, Arabidopsis
keywords: Catalase 3 C-terminal domain; E1 SUMO-activating enzyme (SAE2/SAE1); E2 SUMO-conjugating enzyme; In vitro SUMOylation assay; SUMO; Thioester