Structural analysis of a plant sucrose carrier using monoclonal antibodies and bacteriophage lambda surface display

FEBS Letters

Sauer, Norbert²  Stolz, Jürgen²  Hagemann, Klaus¹  Ludwig, Andreas²  Biesgen, Christian¹  Stadler, Ruth² 

[1] Lehrstuhl für Pflanzenphysiologie, Ruhr-Universität Bochum, Universitätsstraße 150, D-44801 Bochum, Germany;Lehrstuhl Botanik II, Molekulare Pflanzenphysiologie, Universität Erlangen-Nürnberg, Staudtstraße 5, D-91058 Erlangen, Germany

关键词: Monoclonal antibody;    Phage display;    Topology;    Sucrose transport;    Plantago;    CCCP;    carbonyl cyanide-m-chlorophenylhydrazone;    ELISA;    enzyme-linked immunosorbent assay;    FITC;    fluorescenin isothiocyanate;    mAB;    monoclonal antibody;    PAGE;    polyacrylamide gel electrophoresis;    SDS;    sodium dodecyl sulfate;    TRITC;    tetramethylrhodamine B isothiocyanate;   

DOI  :  10.1016/S0014-5793(99)00756-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Monoclonal antibodies were raised and selected against recombinant Plantago major PmSUC2 sucrose carrier protein. Epitopes of two monoclonal antibodies (PS2-1A2 and PS2-4D4) were mapped using N-terminally truncated PmSUC2 proteins and a lambda library displaying random PmSUC2 peptides. PS2-1A2 recognizes an octapeptide close to the N-terminus of PmSUC2, PS2-4D4 binds to a decapeptide at the very C-terminus. Analyses of antibody binding to yeast protoplasts with functionally active, tagged PmSUC2 protein revealed that both epitopes are located in cytoplasmic domains of PmSUC2. These results support a model for plant sucrose transporters containing 12 transmembrane helices with the N-terminus and the C-terminus on the cytoplasmic side of the plasma membrane.

【 授权许可】

Unknown