Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I
Poly(ADP-ribose)polymerase has been purified more than 160,000-fold from Crithidia fasciculata. This is the first PARP isolated to apparent homogeneity from trypanosomatids. The purified enzyme absolutely required DNA for catalytic activity and histones enhanced it 2.5-fold, when the DNA:histone rat...
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2004
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v135_n2_p211_Podesta http://hdl.handle.net/20.500.12110/paper_01666851_v135_n2_p211_Podesta |
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paper:paper_01666851_v135_n2_p211_Podesta2023-06-08T15:15:56Z Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I Crithidia fasciculata D,L-dithiothreitol DNA topoisomerase I DTT NAD+ PARP Poly(ADP-ribose)polymerase TCA Topo Trichloroacetic acid β-Me β-mercaptoethanol β-nicotinamide adenine dinucleotide adenosine DNA topoisomerase hypoxanthine nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine xanthine article catalysis cell nucleus controlled study Crithidia fasciculata enzyme activity enzyme inhibition enzyme isolation enzyme purification gel filtration molecular dynamics nonhuman priority journal Crithidia fasciculata Poly(ADP-ribose)polymerase has been purified more than 160,000-fold from Crithidia fasciculata. This is the first PARP isolated to apparent homogeneity from trypanosomatids. The purified enzyme absolutely required DNA for catalytic activity and histones enhanced it 2.5-fold, when the DNA:histone ratio was 1:1.3. The enzyme required no magnesium or any other metal ion cofactor. The apparent molecular mass of 111kDa, determined by gel filtration would correspond to a dimer of two identical 55-kDa subunits. Activity was inhibited by nicotinamide, 3-aminobenzamide, theophylline, thymidine, xanthine and hypoxanthine but not by adenosine. The enzyme was localized to the cell nucleus. Our findings suggest that covalent poly(ADP-ribosyl)ation of PARP itself or DNA topoisomerase I resulted in the inhibition of their activities and provide an initial biochemical characterization of this covalent post-translational modification in trypanosomatids. © 2004 Elsevier B.V. All rights reserved. 2004 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v135_n2_p211_Podesta http://hdl.handle.net/20.500.12110/paper_01666851_v135_n2_p211_Podesta |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Crithidia fasciculata D,L-dithiothreitol DNA topoisomerase I DTT NAD+ PARP Poly(ADP-ribose)polymerase TCA Topo Trichloroacetic acid β-Me β-mercaptoethanol β-nicotinamide adenine dinucleotide adenosine DNA topoisomerase hypoxanthine nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine xanthine article catalysis cell nucleus controlled study Crithidia fasciculata enzyme activity enzyme inhibition enzyme isolation enzyme purification gel filtration molecular dynamics nonhuman priority journal Crithidia fasciculata |
| spellingShingle |
Crithidia fasciculata D,L-dithiothreitol DNA topoisomerase I DTT NAD+ PARP Poly(ADP-ribose)polymerase TCA Topo Trichloroacetic acid β-Me β-mercaptoethanol β-nicotinamide adenine dinucleotide adenosine DNA topoisomerase hypoxanthine nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine xanthine article catalysis cell nucleus controlled study Crithidia fasciculata enzyme activity enzyme inhibition enzyme isolation enzyme purification gel filtration molecular dynamics nonhuman priority journal Crithidia fasciculata Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I |
| topic_facet |
Crithidia fasciculata D,L-dithiothreitol DNA topoisomerase I DTT NAD+ PARP Poly(ADP-ribose)polymerase TCA Topo Trichloroacetic acid β-Me β-mercaptoethanol β-nicotinamide adenine dinucleotide adenosine DNA topoisomerase hypoxanthine nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine xanthine article catalysis cell nucleus controlled study Crithidia fasciculata enzyme activity enzyme inhibition enzyme isolation enzyme purification gel filtration molecular dynamics nonhuman priority journal Crithidia fasciculata |
| description |
Poly(ADP-ribose)polymerase has been purified more than 160,000-fold from Crithidia fasciculata. This is the first PARP isolated to apparent homogeneity from trypanosomatids. The purified enzyme absolutely required DNA for catalytic activity and histones enhanced it 2.5-fold, when the DNA:histone ratio was 1:1.3. The enzyme required no magnesium or any other metal ion cofactor. The apparent molecular mass of 111kDa, determined by gel filtration would correspond to a dimer of two identical 55-kDa subunits. Activity was inhibited by nicotinamide, 3-aminobenzamide, theophylline, thymidine, xanthine and hypoxanthine but not by adenosine. The enzyme was localized to the cell nucleus. Our findings suggest that covalent poly(ADP-ribosyl)ation of PARP itself or DNA topoisomerase I resulted in the inhibition of their activities and provide an initial biochemical characterization of this covalent post-translational modification in trypanosomatids. © 2004 Elsevier B.V. All rights reserved. |
| title |
Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I |
| title_short |
Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I |
| title_full |
Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I |
| title_fullStr |
Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I |
| title_full_unstemmed |
Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata: Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I |
| title_sort |
purification and properties of poly(adp-ribose)polymerase from crithidia fasciculata: automodification and poly(adp-ribosyl)ation of dna topoisomerase i |
| publishDate |
2004 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v135_n2_p211_Podesta http://hdl.handle.net/20.500.12110/paper_01666851_v135_n2_p211_Podesta |
| _version_ |
1768546015944638464 |