Artículo
Abstract:
The primary structure of several chloroplast fructose-1,6-bisphosphatase (CFBPase) was deduced from DNA sequences, but only spinach, pea and rapeseed enzymes have been characterized structurally. We analyzed whether CFBPases from different phylogenetic origin contain a common epitope. To this end a DNA fragment of 1200 base pairs encoding 338 amino acid residues of wheat CFBPase (38 kDa) was cloned in the expression plasmid pGEX-1 in frame with the gene coding for glutathione S-transferase (GT) of Schistosoma japonicun (26.5 kDa). Upon transformation of Escherichia coli and induction with isopropyl-beta-D-thiogalactopyranoside, centrifugation of the lysate partitioned 10% of the fusion protein in the supernatant fraction and the remaining 90% in the precipitate. The expected 65 kDa protein was purified from both the soluble and the particulate fraction by affinity chromatography on columns of glutathione-agarose. This fusion protein was successfully used to produce a monoclonal antibody that specifically recognized the CFBPase region of the fusion protein but not the GT moiety. Moreover, the monoclonal antibody immunoreacted not only with polypeptides (ca. 40 kDa) present in leaf crude extracts of other varieties of wheat (Triticum spelta, T. aestivum and T. durum), but also with homogeneous preparations of the spinach (Spinacia oleracea) and rapeseed (Brassica napus) enzymes. Thus, the cross reaction of this monoclonal antibody with counterparts from different plant species indicates the persistency of a common epitope through biological evolution.
Registro:
| Documento: | Artículo |
| Título: | Interspecies cross-reactivity of a monoclonal antibody directed against wheat chloroplast fructose-1,6-bisphosphatase. |
| Autor: | Hagelin, K.; Rodriguez-Suarez, R.; Katzen, F.; Wolosiuk, R.A.; Baldi, P.C.; Giambartolomei, G.H.; Fossati, C.A.; Dyer, T. |
| Filiación: | Instituto de Investigaciones Bioquimicas Luis F. Leloir Fundación Campomar, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina |
| Palabras clave: | epitope; fructose bisphosphatase; glutathione transferase; hybrid protein; monoclonal antibody; peptide fragment; amino acid sequence; animal; antibody specificity; article; chloroplast; comparative study; cross reaction; enzymology; Escherichia coli; genetics; immunology; isolation and purification; molecular cloning; molecular evolution; mouse; plant; Schistosoma japonicum; species difference; wheat; Amino Acid Sequence; Animals; Antibodies, Monoclonal; Antibody Specificity; Chloroplasts; Cloning, Molecular; Cross Reactions; Epitopes; Escherichia coli; Evolution, Molecular; Fructose-Bisphosphatase; Glutathione Transferase; Mice; Peptide Fragments; Plants; Recombinant Fusion Proteins; Schistosoma japonicum; Species Specificity; Triticum |
| Año: | 1996 |
| Volumen: | 42 |
| Número: | 5 |
| Página de inicio: | 673 |
| Página de fin: | 682 |
| Título revista: | Cellular and molecular biology (Noisy-le-Grand, France) |
| Título revista abreviado: | Cell. Mol. Biol. (Noisy-le-grand) |
| ISSN: | 01455680 |
| CAS: | fructose bisphosphatase, 9001-52-9; glutathione transferase, 50812-37-8; Antibodies, Monoclonal; Epitopes; Fructose-Bisphosphatase, EC 3.1.3.11; Glutathione Transferase, EC 2.5.1.18; Peptide Fragments; Recombinant Fusion Proteins |
| Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01455680_v42_n5_p673_Hagelin |
Citas:
---------- APA ----------
Hagelin, K., Rodriguez-Suarez, R., Katzen, F., Wolosiuk, R.A., Baldi, P.C., Giambartolomei, G.H., Fossati, C.A.,..., Dyer, T. (1996) . Interspecies cross-reactivity of a monoclonal antibody directed against wheat chloroplast fructose-1,6-bisphosphatase. Cellular and molecular biology (Noisy-le-Grand, France), 42(5), 673-682.Recuperado de https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01455680_v42_n5_p673_Hagelin [ ]
---------- CHICAGO ----------
Hagelin, K., Rodriguez-Suarez, R., Katzen, F., Wolosiuk, R.A., Baldi, P.C., Giambartolomei, G.H., et al. "Interspecies cross-reactivity of a monoclonal antibody directed against wheat chloroplast fructose-1,6-bisphosphatase." Cellular and molecular biology (Noisy-le-Grand, France) 42, no. 5 (1996) : 673-682.Recuperado de https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01455680_v42_n5_p673_Hagelin [ ]
---------- MLA ----------
Hagelin, K., Rodriguez-Suarez, R., Katzen, F., Wolosiuk, R.A., Baldi, P.C., Giambartolomei, G.H., et al. "Interspecies cross-reactivity of a monoclonal antibody directed against wheat chloroplast fructose-1,6-bisphosphatase." Cellular and molecular biology (Noisy-le-Grand, France), vol. 42, no. 5, 1996, pp. 673-682.Recuperado de https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01455680_v42_n5_p673_Hagelin [ ]
---------- VANCOUVER ----------
Hagelin, K., Rodriguez-Suarez, R., Katzen, F., Wolosiuk, R.A., Baldi, P.C., Giambartolomei, G.H., et al. Interspecies cross-reactivity of a monoclonal antibody directed against wheat chloroplast fructose-1,6-bisphosphatase. Cell. Mol. Biol. (Noisy-le-grand). 1996;42(5):673-682.Available from: https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01455680_v42_n5_p673_Hagelin [ ]
