Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase

Photoxidation with methylene blue and rose bengal and chemical modification by diethylpyrocarbonate of pig liver 5-aminolevulinic acid dehydratase produced strong inactivation of the enzyme which was concentration dependent. Loss of enzyme activity by both photoxidation and ethoxyformylation was pH...

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Autores principales:	Fukuda, Haydeé, Paredes, Sergio Raúl, Ferramola, Ana María, Sancovich, Horacio Alberto, Batlle, Alcira María del Carmen
Publicado:	1990
Materias:	5-Aminolevulinic acid dehydratase Diethylpyrocarbonate Histidyl residues Methylene blue Rose Bengal diethyl pyrocarbonate histidine methylene blue porphobilinogen synthase rose bengal animal cell article enzyme active site enzyme modification nonhuman photooxidation swine Animal Binding Sites Enzyme Inhibitors Histidine Hydrogen-Ion Concentration Kinetics Liver Macromolecular Systems Methylene Blue Photochemistry Porphobilinogen Synthase Support, Non-U.S. Gov't Swine Animalia Sus scrofa
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14756366_v3_n4_p295_Fukuda http://hdl.handle.net/20.500.12110/paper_14756366_v3_n4_p295_Fukuda
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_14756366_v3_n4_p295_Fukuda
record_format	dspace
spelling	paper:paper_14756366_v3_n4_p295_Fukuda2023-06-08T16:17:51Z Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase Fukuda, Haydeé Paredes, Sergio Raúl Ferramola, Ana María Sancovich, Horacio Alberto Batlle, Alcira María del Carmen 5-Aminolevulinic acid dehydratase Diethylpyrocarbonate Histidyl residues Methylene blue Rose Bengal diethyl pyrocarbonate histidine methylene blue porphobilinogen synthase rose bengal animal cell article enzyme active site enzyme modification nonhuman photooxidation swine Animal Binding Sites Enzyme Inhibitors Histidine Hydrogen-Ion Concentration Kinetics Liver Macromolecular Systems Methylene Blue Photochemistry Porphobilinogen Synthase Rose Bengal Support, Non-U.S. Gov't Swine Animalia Sus scrofa Photoxidation with methylene blue and rose bengal and chemical modification by diethylpyrocarbonate of pig liver 5-aminolevulinic acid dehydratase produced strong inactivation of the enzyme which was concentration dependent. Loss of enzyme activity by both photoxidation and ethoxyformylation was pH and time-dependent and protected by the presence of the substate and competitive inhibitors. The rate of inactivation was directly related to the state of protonation of histidyl groups, the unprotonated from being modified at a much faster rate than the protonated form. Plots of the pseudo-first order rate constants for 5-aminolevulinic acid dehydratase inactivation against pH resulted in typical titration curves showing inflection points at about pH 6.4 for methylene blue and rose bengal and 6.8 for diethylpyrocarbonate providing further and unequivocal evidence for the existence of critical histidyl groups at the active centre of the enzyme. © 1990 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted. Fil:Fukuda, H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Paredes, S.R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ferramola De Sancovich, A.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Sancovich, H.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Batlle, A.M.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1990 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14756366_v3_n4_p295_Fukuda http://hdl.handle.net/20.500.12110/paper_14756366_v3_n4_p295_Fukuda
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	5-Aminolevulinic acid dehydratase Diethylpyrocarbonate Histidyl residues Methylene blue Rose Bengal diethyl pyrocarbonate histidine methylene blue porphobilinogen synthase rose bengal animal cell article enzyme active site enzyme modification nonhuman photooxidation swine Animal Binding Sites Enzyme Inhibitors Histidine Hydrogen-Ion Concentration Kinetics Liver Macromolecular Systems Methylene Blue Photochemistry Porphobilinogen Synthase Rose Bengal Support, Non-U.S. Gov't Swine Animalia Sus scrofa
spellingShingle	5-Aminolevulinic acid dehydratase Diethylpyrocarbonate Histidyl residues Methylene blue Rose Bengal diethyl pyrocarbonate histidine methylene blue porphobilinogen synthase rose bengal animal cell article enzyme active site enzyme modification nonhuman photooxidation swine Animal Binding Sites Enzyme Inhibitors Histidine Hydrogen-Ion Concentration Kinetics Liver Macromolecular Systems Methylene Blue Photochemistry Porphobilinogen Synthase Rose Bengal Support, Non-U.S. Gov't Swine Animalia Sus scrofa Fukuda, Haydeé Paredes, Sergio Raúl Ferramola, Ana María Sancovich, Horacio Alberto Batlle, Alcira María del Carmen Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
topic_facet	5-Aminolevulinic acid dehydratase Diethylpyrocarbonate Histidyl residues Methylene blue Rose Bengal diethyl pyrocarbonate histidine methylene blue porphobilinogen synthase rose bengal animal cell article enzyme active site enzyme modification nonhuman photooxidation swine Animal Binding Sites Enzyme Inhibitors Histidine Hydrogen-Ion Concentration Kinetics Liver Macromolecular Systems Methylene Blue Photochemistry Porphobilinogen Synthase Rose Bengal Support, Non-U.S. Gov't Swine Animalia Sus scrofa
description	Photoxidation with methylene blue and rose bengal and chemical modification by diethylpyrocarbonate of pig liver 5-aminolevulinic acid dehydratase produced strong inactivation of the enzyme which was concentration dependent. Loss of enzyme activity by both photoxidation and ethoxyformylation was pH and time-dependent and protected by the presence of the substate and competitive inhibitors. The rate of inactivation was directly related to the state of protonation of histidyl groups, the unprotonated from being modified at a much faster rate than the protonated form. Plots of the pseudo-first order rate constants for 5-aminolevulinic acid dehydratase inactivation against pH resulted in typical titration curves showing inflection points at about pH 6.4 for methylene blue and rose bengal and 6.8 for diethylpyrocarbonate providing further and unequivocal evidence for the existence of critical histidyl groups at the active centre of the enzyme. © 1990 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.
author	Fukuda, Haydeé Paredes, Sergio Raúl Ferramola, Ana María Sancovich, Horacio Alberto Batlle, Alcira María del Carmen
author_facet	Fukuda, Haydeé Paredes, Sergio Raúl Ferramola, Ana María Sancovich, Horacio Alberto Batlle, Alcira María del Carmen
author_sort	Fukuda, Haydeé
title	Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
title_short	Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
title_full	Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
title_fullStr	Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
title_full_unstemmed	Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
title_sort	further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase
publishDate	1990
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_14756366_v3_n4_p295_Fukuda http://hdl.handle.net/20.500.12110/paper_14756366_v3_n4_p295_Fukuda
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Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase

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