Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance

Cytochrome P450 hemoproteins (CYPs) are involved in the synthesis of endogenous compounds such as steroids, fatty acids and prostaglandins as well as in the activation and detoxification of foreign compounds including therapeutic drugs. Cytochrome P450 reductase (CPR, E.C.1.6.2.4) transfers electron...

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Publicado:	2008
Materias:	Cytochrome P450 reductase Drug resistance Trypanosoma cruzi amino acid benznidazole cytochrome c cytochrome P450 diphenyliodonium salt flavine adenine nucleotide flavine mononucleotide nifurtimox polyclonal antiserum protein recombinant enzyme reduced nicotinamide adenine dinucleotide phosphate reduced nicotinamide adenine dinucleotide phosphate ferrihemoprotein reductase amino acid sequence amino terminal sequence antibiotic resistance article cell culture clone developmental stage epimastigote Escherichia coli gene sequence microsome molecular weight nonhuman nucleotide sequence priority journal Amino Acid Sequence Animals DNA, Protozoan Drug Resistance Microsomes, Liver Molecular Sequence Data NADPH-Ferrihemoprotein Reductase Rats Recombinant Proteins Transfection Rattus
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v160_n1_p42_Portal http://hdl.handle.net/20.500.12110/paper_01666851_v160_n1_p42_Portal
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_01666851_v160_n1_p42_Portal2023-06-08T15:16:00Z Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance Cytochrome P450 reductase Drug resistance Trypanosoma cruzi amino acid benznidazole cytochrome c cytochrome P450 diphenyliodonium salt flavine adenine nucleotide flavine mononucleotide nifurtimox polyclonal antiserum protein recombinant enzyme reduced nicotinamide adenine dinucleotide phosphate reduced nicotinamide adenine dinucleotide phosphate ferrihemoprotein reductase amino acid sequence amino terminal sequence antibiotic resistance article cell culture clone developmental stage epimastigote Escherichia coli gene sequence microsome molecular weight nonhuman nucleotide sequence priority journal Trypanosoma cruzi Amino Acid Sequence Animals DNA, Protozoan Drug Resistance Escherichia coli Microsomes, Liver Molecular Sequence Data NADPH-Ferrihemoprotein Reductase Rats Recombinant Proteins Transfection Trypanosoma cruzi Escherichia coli Rattus Trypanosoma cruzi Cytochrome P450 hemoproteins (CYPs) are involved in the synthesis of endogenous compounds such as steroids, fatty acids and prostaglandins as well as in the activation and detoxification of foreign compounds including therapeutic drugs. Cytochrome P450 reductase (CPR, E.C.1.6.2.4) transfers electrons from NADPH to a number of hemoproteins such as CYPs, cytochrome c, cytochrome b5, and heme oxygenase. This work presents the complete sequences of three non-allelic CPR genes from Trypanosoma cruzi. The encoded proteins named TcCPR-A, TcCPR-B and TcCPR-C have calculated molecular masses of 68.6 kDa, 78.4 kDa and 71.3 kDa, respectively. Deduced amino acid sequences share 11% amino acid identity, possess the conserved binding domains for FMN, FAD and NADPH and differ in the hydrophobic 27-amino acid residues of the N-terminal extension, which is absent in TcCPR-A. Every T. cruzi CPRs, TcCPR-A, TcCPR-B and TcCPR-C, were cloned and expressed in Escherichia coli. All of the recombinant enzymes reduced cytochrome c in a NADPH absolutely dependent manner with low Km values for this cofactor. They all were also strongly inhibited by diphenyleneiodonium, a classical flavoenzyme inhibitor. In addition, TcCPRs could support CYP activities when assayed in reconstituted systems containing rat liver microsomes. Polyclonal antiserum rose against the recombinant enzymes TcCPR-A and TcCPR-B demonstrated its presence in every T. cruzi developmental stages, with a remarkable expression of TcCPR-A in cell-cultured trypomastigotes. Overexpression of TcCPR-B in T. cruzi epimastigotes increased its resistance to the typical chemotherapeutic agents Nifurtimox and Benznidazole. We suggest a participation of TcCPR-B in the detoxification metabolism of the parasite. © 2008 Elsevier B.V. All rights reserved. 2008 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v160_n1_p42_Portal http://hdl.handle.net/20.500.12110/paper_01666851_v160_n1_p42_Portal
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Cytochrome P450 reductase Drug resistance Trypanosoma cruzi amino acid benznidazole cytochrome c cytochrome P450 diphenyliodonium salt flavine adenine nucleotide flavine mononucleotide nifurtimox polyclonal antiserum protein recombinant enzyme reduced nicotinamide adenine dinucleotide phosphate reduced nicotinamide adenine dinucleotide phosphate ferrihemoprotein reductase amino acid sequence amino terminal sequence antibiotic resistance article cell culture clone developmental stage epimastigote Escherichia coli gene sequence microsome molecular weight nonhuman nucleotide sequence priority journal Trypanosoma cruzi Amino Acid Sequence Animals DNA, Protozoan Drug Resistance Escherichia coli Microsomes, Liver Molecular Sequence Data NADPH-Ferrihemoprotein Reductase Rats Recombinant Proteins Transfection Trypanosoma cruzi Escherichia coli Rattus Trypanosoma cruzi
spellingShingle	Cytochrome P450 reductase Drug resistance Trypanosoma cruzi amino acid benznidazole cytochrome c cytochrome P450 diphenyliodonium salt flavine adenine nucleotide flavine mononucleotide nifurtimox polyclonal antiserum protein recombinant enzyme reduced nicotinamide adenine dinucleotide phosphate reduced nicotinamide adenine dinucleotide phosphate ferrihemoprotein reductase amino acid sequence amino terminal sequence antibiotic resistance article cell culture clone developmental stage epimastigote Escherichia coli gene sequence microsome molecular weight nonhuman nucleotide sequence priority journal Trypanosoma cruzi Amino Acid Sequence Animals DNA, Protozoan Drug Resistance Escherichia coli Microsomes, Liver Molecular Sequence Data NADPH-Ferrihemoprotein Reductase Rats Recombinant Proteins Transfection Trypanosoma cruzi Escherichia coli Rattus Trypanosoma cruzi Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance
topic_facet	Cytochrome P450 reductase Drug resistance Trypanosoma cruzi amino acid benznidazole cytochrome c cytochrome P450 diphenyliodonium salt flavine adenine nucleotide flavine mononucleotide nifurtimox polyclonal antiserum protein recombinant enzyme reduced nicotinamide adenine dinucleotide phosphate reduced nicotinamide adenine dinucleotide phosphate ferrihemoprotein reductase amino acid sequence amino terminal sequence antibiotic resistance article cell culture clone developmental stage epimastigote Escherichia coli gene sequence microsome molecular weight nonhuman nucleotide sequence priority journal Trypanosoma cruzi Amino Acid Sequence Animals DNA, Protozoan Drug Resistance Escherichia coli Microsomes, Liver Molecular Sequence Data NADPH-Ferrihemoprotein Reductase Rats Recombinant Proteins Transfection Trypanosoma cruzi Escherichia coli Rattus Trypanosoma cruzi
description	Cytochrome P450 hemoproteins (CYPs) are involved in the synthesis of endogenous compounds such as steroids, fatty acids and prostaglandins as well as in the activation and detoxification of foreign compounds including therapeutic drugs. Cytochrome P450 reductase (CPR, E.C.1.6.2.4) transfers electrons from NADPH to a number of hemoproteins such as CYPs, cytochrome c, cytochrome b5, and heme oxygenase. This work presents the complete sequences of three non-allelic CPR genes from Trypanosoma cruzi. The encoded proteins named TcCPR-A, TcCPR-B and TcCPR-C have calculated molecular masses of 68.6 kDa, 78.4 kDa and 71.3 kDa, respectively. Deduced amino acid sequences share 11% amino acid identity, possess the conserved binding domains for FMN, FAD and NADPH and differ in the hydrophobic 27-amino acid residues of the N-terminal extension, which is absent in TcCPR-A. Every T. cruzi CPRs, TcCPR-A, TcCPR-B and TcCPR-C, were cloned and expressed in Escherichia coli. All of the recombinant enzymes reduced cytochrome c in a NADPH absolutely dependent manner with low Km values for this cofactor. They all were also strongly inhibited by diphenyleneiodonium, a classical flavoenzyme inhibitor. In addition, TcCPRs could support CYP activities when assayed in reconstituted systems containing rat liver microsomes. Polyclonal antiserum rose against the recombinant enzymes TcCPR-A and TcCPR-B demonstrated its presence in every T. cruzi developmental stages, with a remarkable expression of TcCPR-A in cell-cultured trypomastigotes. Overexpression of TcCPR-B in T. cruzi epimastigotes increased its resistance to the typical chemotherapeutic agents Nifurtimox and Benznidazole. We suggest a participation of TcCPR-B in the detoxification metabolism of the parasite. © 2008 Elsevier B.V. All rights reserved.
title	Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance
title_short	Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance
title_full	Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance
title_fullStr	Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance
title_full_unstemmed	Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance
title_sort	multiple nadph-cytochrome p450 reductases from trypanosoma cruzi. suggested role on drug resistance
publishDate	2008
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v160_n1_p42_Portal http://hdl.handle.net/20.500.12110/paper_01666851_v160_n1_p42_Portal
_version_	1768546016128139264

Multiple NADPH-cytochrome P450 reductases from Trypanosoma cruzi. Suggested role on drug resistance

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