title: Light Regulation of Brassinosteroid Signaling Components: Checking Regulation of Protein Stability in Darkness.
abstract: Environmental conditions can affect stability of proteins at transcriptional or posttranscriptional levels to modulate their functions. Here we describe a method to observe changes in protein stability under different light conditions. In brief, Arabidopsis thaliana seedlings were maintained under various light regimes from continuous light to total darkness or transitions from light to dark, whereafter total protein was extracted from plants. Proteins were measured and resolved on sodium dodecyl sulfate-polyacrylamide gels and transferred to polyvinylidene difluoride membranes. Blots were incubated with the corresponding antibodies for the visualization of protein bands. The protocol described has been successfully applied in wild-type, different transgenic, and mutant background plants to study how light alone or in combination with other factors influences protein stability.
authors.affiliation: School of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul, 151-747, Korea.
authors.forename: Claudia
authors.initials: C
authors.lastname: Corvalán
authors.affiliation: School of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul, 151-747, Korea. shchoe@snu.ac.kr.|Convergence Research Center for Functional Plant Products, Advanced Institutes of Convergence Technology, Gyeonggi-do, 443-270, Korea. shchoe@snu.ac.kr.|Plant Genomics and Breeding Institute, Seoul National University, Seoul, 151-921, Korea. shchoe@snu.ac.kr.
authors.forename: Sunghwa
authors.initials: S
authors.lastname: Choe
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000528:radiation effects*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D060406:Brassinosteroids / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D018506:Gene Expression Regulation, Plant; D059647:Gene-Environment Interaction; D008027:Light; D010507:Periodicity; D010937:Plant Growth Regulators / Q000494:pharmacology*; D030821:Plants, Genetically Modified; D055550:Protein Stability; D036226:Seedlings / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000528:radiation effects*; D015398:Signal Transduction; D013260:Steroids, Heterocyclic / Q000494:pharmacology*
chemical_list: D029681:Arabidopsis Proteins; D060406:Brassinosteroids; D010937:Plant Growth Regulators; D013260:Steroids, Heterocyclic; C023623:brassinolide
keywords: Antibody; Arabidopsis thaliana; Dark treatments; Immunoblot; Light treatments; Protein extraction