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Comparative characterisation of poly(ADP-ribose) polymerase-1 from two mammalian species with different life span

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2000

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Alvarez-Gonzalez, Rafael

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Experimental Gerontology. Elsevier. 2000, 35(8), pp. 989-1002. ISSN 0531-5565. eISSN 1873-6815. Available under: doi: 10.1016/S0531-5565(00)00134-0

Zusammenfassung

DNA damage induced in higher eukaryotes by alkylating agents, oxidants or ionising radiation triggers the synthesis of protein-conjugated poly(ADP-ribose) catalysed by poly(ADP-ribose) polymerase-1 (PARP-1). Previously, cellular poly(ADP-ribosyl)ation capacity has been shown to correlate positively with the life span of mammalian species [Proc. Natl. Acad. Sci. USA 89 (1992) 11 759–11 763]. Here, we have tested whether this correlation results from differences in kinetic parameters of the enzymatic activity of PARP-1. We therefore compared recombinant enzymes, expressed in a baculovirus system, from rat and man as two mammalian species with extremely divergent life span. In standard activity assays performed in the presence of histones as poly(ADP-ribose) acceptors both enzymes showed saturation kinetics with [NAD+]. The kinetic parameters (kcat, km and kcat/km) of the two enzymes were not significantly different. However, in assays assessing the auto-poly(ADP-ribosyl)ation reaction, both enzymes displayed second-order kinetics with respect to [PARP-1], and up to two-fold higher specific activity was observed for human versus rat PARP-1. We conclude that the correlation of poly(ADP-ribosyl)ation capacity with life span is not reflected in the kinetic parameters, but that subtle differences in primary structure of PARP-1 from mammalian species of different longevity may control the extent of the automodification reaction.

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570 Biowissenschaften, Biologie

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Poly(ADP-ribose) polymerase-1, Kinetic parameters, DNA damage, Longevity, Baculovirus

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ISO 690BENEKE, Sascha, Rafael ALVAREZ-GONZALEZ, Alexander BÜRKLE, 2000. Comparative characterisation of poly(ADP-ribose) polymerase-1 from two mammalian species with different life span. In: Experimental Gerontology. Elsevier. 2000, 35(8), pp. 989-1002. ISSN 0531-5565. eISSN 1873-6815. Available under: doi: 10.1016/S0531-5565(00)00134-0
BibTex
@article{Beneke2000Compa-52116,
  year={2000},
  doi={10.1016/S0531-5565(00)00134-0},
  title={Comparative characterisation of poly(ADP-ribose) polymerase-1 from two mammalian species with different life span},
  number={8},
  volume={35},
  issn={0531-5565},
  journal={Experimental Gerontology},
  pages={989--1002},
  author={Beneke, Sascha and Alvarez-Gonzalez, Rafael and Bürkle, Alexander}
}
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