The nodulin VfENOD18 is an ATP-binding protein in infected cells of Vicia faba L. nodules
| Autor: | Becker, JD, Moreira, LM, Kapp, D, Frosch, SC, Pühler, Alfred, Perlick, AM |
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| Jazyk: | angličtina |
| Rok vydání: | 2001 |
| Předmět: |
Methanococcus
Adenosine Triphosphate/metabolism Fabaceae/cytology/genetics/microbiology Blotting Western Molecular Sequence Data Arabidopsis Plant Roots Zea mays Antibodies nitrogen broad bean Adenosine Triphosphate Amino Acid Sequence Microscopy Immunoelectron Symbiosis Plant Proteins Rhizobium leguminosarum Sequence Homology Amino Acid fixation food and beverages Membrane Proteins heterologous expression Fabaceae Recombinant Proteins nitrogen fixation Plant Roots/genetics/microbiology/ultrastructure Soybeans Plant Proteins/genetics/immunology/metabolism MJ0577 superfamily nodule-specific gene expression Protein Binding |
| Zdroj: | Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP |
| Popis: | Recently we described the novel nodulin gene VfENOD18, whose corresponding transcripts were restricted to the nitrogen-fixing zone III of broad bean root nodules. To characterize VfENOD18 on the protein level, polyclonal antibodies were generated using the purified recombinant VfENOD18 protein produced in Escherichia coli by employing the pMAL-c expression system. These antibodies recognized immunoreactive proteins isolated from indeterminate nodules of different leguminous plants, but also from non-symbiotic tissues of Glycine max and from tissues of Arabidopsis thaliana and Zea mays. Using immunogold labelling the nodulin VfENOD18 was localized to the cytoplasm of infected cells in the nitrogen-fixing zone of broad bean nodules. Due to the homology of the VfENOD18 sequence to that of the ATP-binding protein MJ0577 from the hyperthermophile Methanococcus jannaschii the recombinant VfENOD18 protein was tested for ATP-binding. Using the biotin photoaffinity ATP analogue 8N(3)ATP[gamma ]biotin it could be demonstrated that VfENOD18 is an ATP-binding protein. PCR experiments revealed that the amino acid sequences of the putative C-terminal ATP-binding sites of the VfENOD18 homologues from Lens culinaris, Vicia hirsuta, Vicia sativa and Vicia villosa were conserved. We propose that VfENOD18 is a member of a novel family of ATP-binding proteins in plants. |
| Databáze: | OpenAIRE |
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