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

FEBS Letters

Desikan, Radhika²  Rock, Christopher D.¹  Neill, Steven J.²  Hagenbeek, Dik¹ 

[1] Department of Biology, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China;Department of Biological and Biomedical Sciences, University of the West of England, Bristol, Coldharbour Lane, Bristol, BS16 1QY, UK

关键词: Abscisic acid;    Biosensor;    Flow cytometry;    Glycoprotein;    Green fluorescent protein;    Plasma membrane;    Surface plasmon resonance;    ABA;    abscisic acid;    FITC;    fluorescein isothiocyanate;    GFP;    green fluorescent protein;    GP;    glycoprotein;    GUS;    β-glucuronidase;    LUC;    firefly luciferase;    PBS;    phosphate-buffered saline;    PM;    plasma membrane;    SPR;    surface plasmon resonance;    w/v;    weight/volume;   

DOI  :  10.1016/S0014-5793(99)00972-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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, β-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.

【 授权许可】

Unknown