Flow cytometry and surface plasmon resonance analyses demonstrate that the monoclonal antibody JIM19 interacts with a rice cell surface component involved in abscisic acid signalling in protoplasts.
Abscisic acid (ABA) is a plant hormone involved in many developmental and physiological processes, but as yet, no ABA receptor has been identified. Flow cytometry of rice protoplasts and immunoblotting of purified plasma membranes (PMs) have been used to demonstrate that the monoclonal antibody JIM19 recognizes carbohydrate epitopes of cell surface glycoproteins. Using surface plasmon resonance technology specific binding of PMs to JIM19 was observed. Such interaction was antagonized significantly by ABA, but not by the biologically inactive ABA catabolite phaseic acid. These in vitro interactions were correlated with the biological activities of JIM19, ABA and phaseic acid on activation of the ABA-inducible Em promoter using two different transient reporter gene assays, beta-glucuronidase/luciferase and quantitative flow cytometry of Aequoria green fluorescent protein. Pre-treatment with JIM19 resulted in significant inhibition of ABA-inducible gene expression. Taken together, these data suggest that JIM19 interacts with a functional PM complex involved in ABA signalling.
Desikan
R
R
Department of Biological and Biomedical Sciences, University of the West of England, Bristol, UK.
Hagenbeek
D
D
Neill
S J
SJ
Rock
C D
CD
D000040:Abscisic Acid / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D000939:Epitopes; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D008562:Membrane Glycoproteins / Q000276:immunology; D012275:Oryza / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D010937:Plant Growth Regulators / Q000378:metabolism*; D010940:Plant Proteins / Q000276:immunology; D011523:Protoplasts / Q000378:metabolism; D015398:Signal Transduction; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008562:Membrane Glycoproteins; D010937:Plant Growth Regulators; D010940:Plant Proteins; D000040:Abscisic Acid