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Abstract:

1. 1. A simpler method for purifying human red cell deaminase, using a mixture of n-butanol and chloroform, which denatures hemoglobin, followed by ammonium sulphate fractionation, heat treatment. Sephadex G-100 and DEAE-cellulose chromatography, yielding a 3400 fold purified enzyme is described. 2. 2. Some properties of purified deaminase were studied. The enzyme seems to have a strict requirement for oxygen, neither PBG consumption nor uroporphyrinogens formation were measured under anaerobiosis. 3. 3. Uroporphyrinogens formation was linear with both protein and time over a wide range of enzyme concentration and up to 2 h. 4. 4. The optimum pH was 7.4 and the mol. wt was 40,000 ± 4000. The enzyme was heat-stable and increased its activity by heating. 5. 5. Ammonium and hydroxylamine ions inhibited the reaction. K+ and Na+ ions did not greatly affect activity, while most divalent cations tested significantly diminished uroporphyrinogen formation and to a lesser degree PBG consumption. 6. 6. Direct plots of velocity against PBG concentration were hyperbolic, however double-reciprocal plots were non-linear. Hill plots gave an n value of 2 and Eadie plots were bell-shaped, indicating the existence of weakly positive cooperative effect between 2 binding sites for PBG per molecule of deaminase. © 1985.

Registro:

Documento: Artículo
Título:Human red cell porphobilinogen deaminase. a simpler method of purification and some unusual properties
Autor:Fumagalli, S.A.; Kotler, M.L.; Rossetti, M.V.; Del C. Batlle, A.M.
Filiación:Centro de Investigaciones sobre Porfirinas y Porfirias, CIPYP, (CONICET y FCE y N. UBA), Ciudad Universitaria, Pabellon II, 4/dg piso, 1428 Buenos Aires, Argentina
Palabras clave:porphobilinogen deaminase; uroporphyrinogen; blood and hemopoietic system; erythrocyte; human; human cell; 1-Butanol; Ammonia-Lyases; Ammonium Sulfate; Butanols; Chloroform; Chromatography; Drug Stability; Erythrocytes; Human; Hydrogen-Ion Concentration; Hydroxymethylbilane Synthase; Kinetics; Magnesium; Magnesium Chloride; Molecular Weight; Oxygen; Sodium Chloride; Support, Non-U.S. Gov't; Temperature; Uroporphyrinogens
Año:1985
Volumen:17
Número:4
Página de inicio:485
Página de fin:494
DOI: http://dx.doi.org/10.1016/0020-711X(85)90144-2
Título revista:International Journal of Biochemistry
Título revista abreviado:Int. J. Biochem.
ISSN:0020711X
CODEN:IJBOB
CAS:porphobilinogen deaminase, 9036-47-9, 9074-91-3; uroporphyrinogen, 35465-57-7; 1-Butanol, 71-36-3; Ammonia-Lyases, EC 4.3.1.; Ammonium Sulfate, 7783-20-2; Butanols; Chloroform, 67-66-3; Hydroxymethylbilane Synthase, EC 4.3.1.8; Magnesium Chloride, 7786-30-3; Magnesium, 7439-95-4; Oxygen, 7782-44-7; Sodium Chloride, 7647-14-5; Uroporphyrinogens
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0020711X_v17_n4_p485_Fumagalli

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Citas:

---------- APA ----------
Fumagalli, S.A., Kotler, M.L., Rossetti, M.V. & Del C. Batlle, A.M. (1985) . Human red cell porphobilinogen deaminase. a simpler method of purification and some unusual properties. International Journal of Biochemistry, 17(4), 485-494.
http://dx.doi.org/10.1016/0020-711X(85)90144-2
---------- CHICAGO ----------
Fumagalli, S.A., Kotler, M.L., Rossetti, M.V., Del C. Batlle, A.M. "Human red cell porphobilinogen deaminase. a simpler method of purification and some unusual properties" . International Journal of Biochemistry 17, no. 4 (1985) : 485-494.
http://dx.doi.org/10.1016/0020-711X(85)90144-2
---------- MLA ----------
Fumagalli, S.A., Kotler, M.L., Rossetti, M.V., Del C. Batlle, A.M. "Human red cell porphobilinogen deaminase. a simpler method of purification and some unusual properties" . International Journal of Biochemistry, vol. 17, no. 4, 1985, pp. 485-494.
http://dx.doi.org/10.1016/0020-711X(85)90144-2
---------- VANCOUVER ----------
Fumagalli, S.A., Kotler, M.L., Rossetti, M.V., Del C. Batlle, A.M. Human red cell porphobilinogen deaminase. a simpler method of purification and some unusual properties. Int. J. Biochem. 1985;17(4):485-494.
http://dx.doi.org/10.1016/0020-711X(85)90144-2