Abstract:
1. 1. Evidence for dissociation, renaturation, re-association and re-hybridization of bovine liver aminolaevulinate dehydratase attached to Sepharose 4B is reported. 2. 2. When insolubilized enzyme was treated with 3 and 6 M urea, non covalently bound subunits were dissociated and detected in the eluate; these subunits can be re-associated into a soluble functioning enzyme with a specific activity close to that of the original pure soluble dehydratase preparation. 3. 3. After being washed with a renaturing buffer mixture, the matrix-bound subunits recovered a level of enzymatic activity equal to 50 and 20% of that of the immobilized native aminolaevulinate dehydratase. 4. 4. The reversibility of the dissociation process was investigated. Bound-subunits dehydratase can associate with nascent soluble bovine liver aminolaevulinate dehydratase subunits in situ. The product of such treatment, bound-re-associated enzyme, has the same activity as that of the original bound-dehydratase. The matrix-bound-dissociated bovine liver enzyme was also re-hybridized with soluble dehydratase subunits from E. gracilis. 5. 5. The apparent Km and optimum pH of the immobilized subunits were the same as those of the bound-octameric enzyme. 6. 6. A scheme is proposed, explaining the sequence of reactions leading from the bound-octameric dehydratase to the possible different derivatives, formed during the dissociation and re-association experiments. © 1978.
Registro:
Documento: |
Artículo
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Título: | Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity |
Autor: | Del C. Batlle, A.M.; Stella, A.M.; Ferramola, A.M.; Sopena, Y.; De Xifra, E.A.W.; Sancovich, H.A. |
Filiación: | Centro de Investigaciones sobre Porfirinas y Porfirias. Departamento de Quimica Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Nunez, 1428 Buenos Aires, Argentina
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Palabras clave: | enzyme; porphobilinogen synthase; porphyrin; animal experiment; cattle; liver; Animals; Cattle; Chemistry; Enzymes, Immobilized; Hydrogen-Ion Concentration; Kinetics; Liver; Macromolecular Substances; Models, Chemical; Porphobilinogen Synthase; Protein Denaturation; Sepharose; Solubility |
Año: | 1978
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Volumen: | 9
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Número: | 6
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Página de inicio: | 401
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Página de fin: | 406
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DOI: |
http://dx.doi.org/10.1016/0020-711X(78)90053-8 |
Título revista: | International Journal of Biochemistry
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Título revista abreviado: | Int. J. Biochem.
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ISSN: | 0020711X
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CODEN: | IJBOB
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CAS: | porphobilinogen synthase, 9036-37-7; porphyrin, 24869-67-8; Enzymes, Immobilized; Macromolecular Substances; Porphobilinogen Synthase, 4.2.1.24; Sepharose, 9012-36-6
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PDF: | https://bibliotecadigital.exactas.uba.ar/download/paper/paper_0020711X_v9_n6_p401_DelCBatlle.pdf |
Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0020711X_v9_n6_p401_DelCBatlle |
Referencias:
- Batlle, Ferramola, Grinstein, Purification and general properties of delta aminolaevulic dehydratase from cow liver (1967) Biochem. J., 104, pp. 244-249
- Batlle, Ferramola, Grinstein, Delta aminolaevulinate dehydratase (cow liver) (1970) Methods in Enzymology, 17, pp. 216-220. , H. Tabor, C. Tabor, Academic Press, Part A
- Bruch, Schnackerz, Gracy, Matrix-bound phosphoglucose isomerase-formation and properties of monomers and hybrids (1976) Eur. J. Biochem., 68, pp. 153-158
- Carvajal, Martinez, Fernandez, Immobilized monomers of human liver arginase (1977) Biochim. biophys. Acta, 481, pp. 177-183
- Chan, Matrix-bound protein subunits (1970) Biochim. biophys. Acta, 41, pp. 1198-1204
- Chan, Active subunits of trans-aldolase bound to Sepharose (1973) Ew. J. Biochem., 40, pp. 533-541
- Chan, Mawer, Studies on protein subunits II—preparation and properties of active subunits of aldolasc bound to a matrix (1972) Archs Biochem. Biophys., 149, pp. 136-145
- Feldman, Zfisel, Helmrfioi, Complementation of subunits from glycogen phosphorylases of frog and rabbit skeletal muscle and rabbit liver (1976) European Journal of Biochemistry, 65, pp. 285-291
- Klotz, Lanzfrman, Darnell, Quaternary structure of proteins (1970) A. Rev. Biochem., 39, pp. 25-62
- Nagradova, Golovina, Mevkh, Immobilized dimers of d-glyceraldchyde-3-phosphate dehydrogenase (1974) FEBS Lett., 49, pp. 242-245
- Nandi, Inhibition of delta aminolevulinic acid dehydratase of Rhodopseudomonas spheroides by urea guanidine and methyl derivatives of urea (1971) Archives of Biochemistry and Biophysics, 142, p. 157 162
- Nandi, Bakfr-Cohen, Shkmin, Delta aminolevulinic acid dehydratase of Rh. spheroicles. I. Isolation and mechanism (1968) J. biol. Chem., 243, pp. 1224-1230
- Shemin, 5-Aminolaevulinic acid dehydratase: structure, function and mechanism (1976) Phil. Trans. R. Soc. B., 273, pp. 109-115
- Sthlla, Batlle, Porphyrin biosynthesis-immobilized enzymes and ligands V Purification of aminolaevulinate dehydratase from bovine liver by affinity chromatoeraphy (1977) International Journal of Biochemistry, 8, pp. 353-358
- Stella, Battle, Porphyrin biosynthesis immobilized enzymes and ligands VIII Studies on the purification of delta aminolaevulinate dehydratase from Euglena gracilis (1978) Pl. Sci. Lett., , In press
- Sthlla, Wider De Xifra, Baille, Porphyrin biosynthesis—immobilized enzymes and ligands IV Studies on aminolaevulinate dehydratase attached to Sepharose (1977) Molec. Cell. Biochem., 16, p. 97 105
- Weber, Osborn, The reliability of molecular weight determination by dodecil sulphate in polyacrylamidc gel-electrophoresis (1969) J. biol. Chem., 244, pp. 4406-4412
Citas:
---------- APA ----------
Del C. Batlle, A.M., Stella, A.M., Ferramola, A.M., Sopena, Y., De Xifra, E.A.W. & Sancovich, H.A.
(1978)
. Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity. International Journal of Biochemistry, 9(6), 401-406.
http://dx.doi.org/10.1016/0020-711X(78)90053-8---------- CHICAGO ----------
Del C. Batlle, A.M., Stella, A.M., Ferramola, A.M., Sopena, Y., De Xifra, E.A.W., Sancovich, H.A.
"Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity"
. International Journal of Biochemistry 9, no. 6
(1978) : 401-406.
http://dx.doi.org/10.1016/0020-711X(78)90053-8---------- MLA ----------
Del C. Batlle, A.M., Stella, A.M., Ferramola, A.M., Sopena, Y., De Xifra, E.A.W., Sancovich, H.A.
"Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity"
. International Journal of Biochemistry, vol. 9, no. 6, 1978, pp. 401-406.
http://dx.doi.org/10.1016/0020-711X(78)90053-8---------- VANCOUVER ----------
Del C. Batlle, A.M., Stella, A.M., Ferramola, A.M., Sopena, Y., De Xifra, E.A.W., Sancovich, H.A. Porphyrin biosynthesis-immobilized enzymes and ligands-X. A novel approach to the study of the relationship between the quaternary structure of aminolaevulinate dehydratase and its activity. Int. J. Biochem. 1978;9(6):401-406.
http://dx.doi.org/10.1016/0020-711X(78)90053-8