Single Amino Acid Substitution Enhances Bacterial Expression of PARP-4D214A

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 2003 Mar 7

PMID 12619884

Citations 6

Authors

Steve N Gagnon

Serge Desnoyers

Affiliations

Soon will be listed here.

Abstract

Poly(ADP-ribose) polymerase-1 (PARP-1) is the canonical member of the PARP family of enzymes and modulates many crucial nuclear functions. PARP-1 is involved in apoptosis and is the substrate of caspase-3, a protease that cleaves PARP-1 at the conserved sequence 211DEVD214. To generate a caspase-3-uncleavable PARP-1, we introduced an amino acid substitution D214-->A214 at the site of cleavage. We observed that following over-expression in bacteria, the mutant protein HIS-PARP-1D214A was expressed several-fold more than a unmutated copy, HIS-PARP-1. The specific activity of HIS-PARP-1 enzyme in total bacterial extracts was 6.94 U/mg and 4.61 U/mg for HIS-PARP-1D214A. This approach should provide new avenues for crystallographic study of PARP-1 as well as new information for drug design targeting PARP-1.

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References

Mazen A, Molinete M, Simonin F, Gradwohl G, Poirier G, de Murcia G . Poly(ADP-ribose)polymerase: a novel finger protein. Nucleic Acids Res. 1989; 17(12):4689-98. PMC: 318025. DOI: 10.1093/nar/17.12.4689. View

Smulson M, Pang D, Jung M, Dimtchev A, Chasovskikh S, Spoonde A . Irreversible binding of poly(ADP)ribose polymerase cleavage product to DNA ends revealed by atomic force microscopy: possible role in apoptosis. Cancer Res. 1998; 58(16):3495-8. View

Zahradka P, Ebisuzaki K . Poly(ADP-ribose) polymerase is a zinc metalloenzyme. Eur J Biochem. 1984; 142(3):503-9. DOI: 10.1111/j.1432-1033.1984.tb08314.x. View

Ruf A, Mennissier de Murcia J, de Murcia G, Schulz G . Structure of the catalytic fragment of poly(AD-ribose) polymerase from chicken. Proc Natl Acad Sci U S A. 1996; 93(15):7481-5. PMC: 38770. DOI: 10.1073/pnas.93.15.7481. View

Ame J, Rolli V, Schreiber V, Niedergang C, Apiou F, Decker P . PARP-2, A novel mammalian DNA damage-dependent poly(ADP-ribose) polymerase. J Biol Chem. 1999; 274(25):17860-8. DOI: 10.1074/jbc.274.25.17860. View

Wang Z, Auer B, Stingl L, Berghammer H, Haidacher D, Schweiger M . Mice lacking ADPRT and poly(ADP-ribosyl)ation develop normally but are susceptible to skin disease. Genes Dev. 1995; 9(5):509-20. DOI: 10.1101/gad.9.5.509. View

Kickhoefer V, Siva A, Kedersha N, Inman E, Ruland C, Streuli M . The 193-kD vault protein, VPARP, is a novel poly(ADP-ribose) polymerase. J Cell Biol. 1999; 146(5):917-28. PMC: 2169495. DOI: 10.1083/jcb.146.5.917. View

Vuillard L, Rabilloud T . Non-detergent sulphobetaines: a new class of mild solubilization agents for protein purification. Biochem J. 1995; 305 ( Pt 1):337-43. PMC: 1136468. DOI: 10.1042/bj3050337. View

Giner H, Simonin F, de Murcia G . Overproduction and large-scale purification of the human poly(ADP-ribose) polymerase using a baculovirus expression system. Gene. 1992; 114(2):279-83. DOI: 10.1016/0378-1119(92)90588-g. View

10.

Wielckens K, Bredehorst R, Hilz H . Quantification of protein-bound ADP-ribosyl and (ADP-ribosyl)n residues. Methods Enzymol. 1984; 106:472-82. DOI: 10.1016/0076-6879(84)06051-1. View

11.

Molinete M, Gradwohl G, Simonin F, de Murcia G . Zinc-binding domain of poly(ADP-ribose)polymerase participates in the recognition of single strand breaks on DNA. J Mol Biol. 1989; 210(1):229-33. DOI: 10.1016/0022-2836(89)90302-1. View

12.

Smith S, Giriat I, Schmitt A, de Lange T . Tankyrase, a poly(ADP-ribose) polymerase at human telomeres. Science. 1998; 282(5393):1484-7. DOI: 10.1126/science.282.5393.1484. View

13.

de Murcia G, Menissier de Murcia J . Poly(ADP-ribose) polymerase: a molecular nick-sensor. Trends Biochem Sci. 1994; 19(4):172-6. DOI: 10.1016/0968-0004(94)90280-1. View

14.

Knight M, Chambers P . Production, extraction, and purification of human poly(ADP-ribose) polymerase-1 (PARP-1) with high specific activity. Protein Expr Purif. 2001; 23(3):453-8. DOI: 10.1006/prep.2001.1513. View

15.

Ha H, Snyder S . Poly(ADP-ribose) polymerase is a mediator of necrotic cell death by ATP depletion. Proc Natl Acad Sci U S A. 1999; 96(24):13978-82. PMC: 24176. DOI: 10.1073/pnas.96.24.13978. View

16.

Shah G, Poirier D, Duchaine C, Brochu G, Desnoyers S, Lagueux J . Methods for biochemical study of poly(ADP-ribose) metabolism in vitro and in vivo. Anal Biochem. 1995; 227(1):1-13. DOI: 10.1006/abio.1995.1245. View

17.

Johansson M . A human poly(ADP-ribose) polymerase gene family (ADPRTL): cDNA cloning of two novel poly(ADP-ribose) polymerase homologues. Genomics. 1999; 57(3):442-5. DOI: 10.1006/geno.1999.5799. View

18.

Masutani M, Suzuki H, Kamada N, Watanabe M, Ueda O, Nozaki T . Poly(ADP-ribose) polymerase gene disruption conferred mice resistant to streptozotocin-induced diabetes. Proc Natl Acad Sci U S A. 1999; 96(5):2301-4. PMC: 26778. DOI: 10.1073/pnas.96.5.2301. View

19.

Gradwohl G, Menissier de Murcia J, Molinete M, Simonin F, Koken M, Hoeijmakers J . The second zinc-finger domain of poly(ADP-ribose) polymerase determines specificity for single-stranded breaks in DNA. Proc Natl Acad Sci U S A. 1990; 87(8):2990-4. PMC: 53819. DOI: 10.1073/pnas.87.8.2990. View

20.

Althaus F, Richter C . ADP-ribosylation of proteins. Enzymology and biological significance. Mol Biol Biochem Biophys. 1987; 37:1-237. View