title: Detection and Quantification of Protein Aggregates in Plants.
abstract: Plants are constantly exposed to a complex and changing environment that challenges their cellular homeostasis. Stress responses triggered as a consequence of unfavorable conditions result in increased protein aggregate formation at the cellular level. When the formation of misfolded proteins surpasses the capacity of the cell to remove them, insoluble protein aggregates accumulate. In the animal field, an enormous effort is being placed to uncover the mechanisms regulating aggregate formation because of its implications in many important human diseases. Because of its importance for cellular functionality and fitness, it is equally important to expand plant research in this field. Here, we describe a cell fractionation-based method to obtain very pure insoluble protein aggregate fractions that can be subsequently semiquantified using image analysis. This method can be used as a first step to evaluate whether a particular condition results in an alteration of protein aggregate formation levels.
authors.affiliation: Centre for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), 08195, Bellaterra, Spain.
authors.forename: Marc
authors.initials: M
authors.lastname: Planas-Marquès
authors.affiliation: Centre for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), 08195, Bellaterra, Spain.
authors.forename: Saul
authors.initials: S
authors.lastname: Lema A
authors.affiliation: Centre for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), 08195, Bellaterra, Spain. nuria.sanchez-coll@cragenomica.es.
authors.forename: Núria S
authors.initials: NS
authors.lastname: Coll
mesh_terms: D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D010944:Plants / Q000737:chemistry / Q000235:genetics; D066329:Protein Aggregates / Q000235:genetics*; D014176:Protein Biosynthesis / Q000235:genetics
chemical_list: D010940:Plant Proteins; D066329:Protein Aggregates
keywords: Immunoblot; Plants; Protein aggregates; Protein fractionation; Proteostasis; Protocols; Silver stain