title: Screening for inhibitors of plant protease D1 using novel monoclonal antibodies directed against its carboxyl terminal.
abstract: Carboxyl terminal processing protease of D1 protein (CtpA) catalyzes carboxyl terminal processing of D1 protein, which is predicted to be an excellent target for a general broad-spectrum herbicide. In this study, the CtpA gene from spinach cDNA was cloned and overexpressed and the recombinant CtpA fusion protein (rCtpA) was used as antigen to immunize BALB/c mice for the production of monoclonal antibody (MAb). Western blot and ELISA results indicated that both rCtpA and the PDZ domain protein of CtpA had specific binding abilities to MAbs, while the specificity and sensitivity of rCtpA were much higher than that of the PDZ domain. These results suggest that parts of the antigen determinant of CtpA were located in the PDZ domain. The MAbs and related results obtained in this study proved the feasibility of high-throughput screening of lead compounds for protease as inhibitors and mechanism analysis of CtpA enzyme.
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Wuhan, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.forename: Weiguo
authors.initials: W
authors.lastname: Li
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Lu
authors.initials: L
authors.lastname: Feng
authors.forename: Yanli
authors.initials: Y
authors.lastname: Liu
authors.forename: Chao
authors.initials: C
authors.lastname: Qi
mesh_terms: D020418:Algal Proteins; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism*; D015153:Blotting, Western; D002268:Carboxypeptidases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D051379:Mice; D008807:Mice, Inbred BALB C; D054731:PDZ Domains / Q000235:genetics / Q000502:physiology*; D043484:Proprotein Convertases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D018724:Spinacia oleracea / Q000235:genetics* / Q000378:metabolism
chemical_list: D020418:Algal Proteins; D000911:Antibodies, Monoclonal; D002268:Carboxypeptidases; D043484:Proprotein Convertases; C073491:photosystem II, D1 polypeptide carboxy-terminal processing protease