Characterization of porphobilinogen deaminase from rat liver

Porphobilinogen deaminase (porphobilinogen ammonia-lyase, EC 4.3.1.8) was isolated from rat liver. The final preparation was homogeneous according to polyacrylamide gel electrophoresis and immunodiffusion criteria. Electrophoresis of the native enzyme revealed a single band of activity which was dis...

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Detalles Bibliográficos
Autores principales: Mazzetti, Marta Blanca, Tomio, Josefina María
Publicado: 1988
Materias:
(Rat liver)
Enzyme purification
Heme synthesis
Porphobilinogen deaminase characterization
liver enzyme
porphobilinogen deaminase
animal cell
electrophoresis
nonhuman
priority journal
rat
Amino Acids
Ammonia-Lyases
Animal
Hydrogen-Ion Concentration
Hydroxymethylbilane Synthase
Isoelectric Point
Kinetics
Liver
Macromolecular Systems
Molecular Weight
Protein Denaturation
Rats
Spectrum Analysis
Sulfhydryl Reagents
Support, Non-U.S. Gov't
Temperature
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674838_v957_n1_p97_Mazzetti
http://hdl.handle.net/20.500.12110/paper_01674838_v957_n1_p97_Mazzetti
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paper:paper_01674838_v957_n1_p97_Mazzetti
record_format dspace
spelling paper:paper_01674838_v957_n1_p97_Mazzetti2023-06-08T15:16:25Z Characterization of porphobilinogen deaminase from rat liver Mazzetti, Marta Blanca Tomio, Josefina María (Rat liver) Enzyme purification Heme synthesis Porphobilinogen deaminase characterization liver enzyme porphobilinogen deaminase animal cell electrophoresis nonhuman priority journal rat Amino Acids Ammonia-Lyases Animal Hydrogen-Ion Concentration Hydroxymethylbilane Synthase Isoelectric Point Kinetics Liver Macromolecular Systems Molecular Weight Protein Denaturation Rats Spectrum Analysis Sulfhydryl Reagents Support, Non-U.S. Gov't Temperature Porphobilinogen deaminase (porphobilinogen ammonia-lyase, EC 4.3.1.8) was isolated from rat liver. The final preparation was homogeneous according to polyacrylamide gel electrophoresis and immunodiffusion criteria. Electrophoresis of the native enzyme revealed a single band of activity which was distributed into three bands after incubation with porphobilinogen. When electrophoresed under denaturing condition it displayed a single polypeptide band with a molecular weight of 42 000 confirmed by exclusion chromatography and by sucrose density gradient centrifugattion. The enzyme showed a pH optimum of 7.5 both in 0.1 M sodium phosphate and 0.05 M Tris-HCl buffer, when assayed at 37°C. An isoelectric point of 4.9 for the native purified protein was found. Hepatic porphobilinogen deaminase was remarkably heat-stable showing maximum activity at 55-60°C with one break in the Arrhenius plot. The kinetic behaviour of the purified enzyme followed the typical Michaelis-Menten kinetics with values of Km = 17 μM and Vmax = 29.4 units power mg in 0.1 M phosphate buffer at 37°C. The amino acid composition was determined, showing that the enzyme had a low content of sulphur-containing amino acids and a considerable number of acidic residues per mol of polypeptide chain. Reagents known to interact with sulphydryl groups have small effect on rat liver enzyme activity. © 1988. Fil:Mazzetti, M.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Tomio, J.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1988 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674838_v957_n1_p97_Mazzetti http://hdl.handle.net/20.500.12110/paper_01674838_v957_n1_p97_Mazzetti
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic (Rat liver)
Enzyme purification
Heme synthesis
Porphobilinogen deaminase characterization
liver enzyme
porphobilinogen deaminase
animal cell
electrophoresis
nonhuman
priority journal
rat
Amino Acids
Ammonia-Lyases
Animal
Hydrogen-Ion Concentration
Hydroxymethylbilane Synthase
Isoelectric Point
Kinetics
Liver
Macromolecular Systems
Molecular Weight
Protein Denaturation
Rats
Spectrum Analysis
Sulfhydryl Reagents
Support, Non-U.S. Gov't
Temperature
spellingShingle (Rat liver)
Enzyme purification
Heme synthesis
Porphobilinogen deaminase characterization
liver enzyme
porphobilinogen deaminase
animal cell
electrophoresis
nonhuman
priority journal
rat
Amino Acids
Ammonia-Lyases
Animal
Hydrogen-Ion Concentration
Hydroxymethylbilane Synthase
Isoelectric Point
Kinetics
Liver
Macromolecular Systems
Molecular Weight
Protein Denaturation
Rats
Spectrum Analysis
Sulfhydryl Reagents
Support, Non-U.S. Gov't
Temperature
Mazzetti, Marta Blanca
Tomio, Josefina María
Characterization of porphobilinogen deaminase from rat liver
topic_facet (Rat liver)
Enzyme purification
Heme synthesis
Porphobilinogen deaminase characterization
liver enzyme
porphobilinogen deaminase
animal cell
electrophoresis
nonhuman
priority journal
rat
Amino Acids
Ammonia-Lyases
Animal
Hydrogen-Ion Concentration
Hydroxymethylbilane Synthase
Isoelectric Point
Kinetics
Liver
Macromolecular Systems
Molecular Weight
Protein Denaturation
Rats
Spectrum Analysis
Sulfhydryl Reagents
Support, Non-U.S. Gov't
Temperature
description Porphobilinogen deaminase (porphobilinogen ammonia-lyase, EC 4.3.1.8) was isolated from rat liver. The final preparation was homogeneous according to polyacrylamide gel electrophoresis and immunodiffusion criteria. Electrophoresis of the native enzyme revealed a single band of activity which was distributed into three bands after incubation with porphobilinogen. When electrophoresed under denaturing condition it displayed a single polypeptide band with a molecular weight of 42 000 confirmed by exclusion chromatography and by sucrose density gradient centrifugattion. The enzyme showed a pH optimum of 7.5 both in 0.1 M sodium phosphate and 0.05 M Tris-HCl buffer, when assayed at 37°C. An isoelectric point of 4.9 for the native purified protein was found. Hepatic porphobilinogen deaminase was remarkably heat-stable showing maximum activity at 55-60°C with one break in the Arrhenius plot. The kinetic behaviour of the purified enzyme followed the typical Michaelis-Menten kinetics with values of Km = 17 μM and Vmax = 29.4 units power mg in 0.1 M phosphate buffer at 37°C. The amino acid composition was determined, showing that the enzyme had a low content of sulphur-containing amino acids and a considerable number of acidic residues per mol of polypeptide chain. Reagents known to interact with sulphydryl groups have small effect on rat liver enzyme activity. © 1988.
author Mazzetti, Marta Blanca
Tomio, Josefina María
author_facet Mazzetti, Marta Blanca
Tomio, Josefina María
author_sort Mazzetti, Marta Blanca
title Characterization of porphobilinogen deaminase from rat liver
title_short Characterization of porphobilinogen deaminase from rat liver
title_full Characterization of porphobilinogen deaminase from rat liver
title_fullStr Characterization of porphobilinogen deaminase from rat liver
title_full_unstemmed Characterization of porphobilinogen deaminase from rat liver
title_sort characterization of porphobilinogen deaminase from rat liver
publishDate 1988
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01674838_v957_n1_p97_Mazzetti
http://hdl.handle.net/20.500.12110/paper_01674838_v957_n1_p97_Mazzetti
work_keys_str_mv AT mazzettimartablanca characterizationofporphobilinogendeaminasefromratliver
AT tomiojosefinamaria characterizationofporphobilinogendeaminasefromratliver
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