细胞技术

A general method to express and purify full-length human poly(ADP-ribose) polymerase-1 (PARP-1), individual PARP-1 domains, and groups of PARP-1 domains from Escherichia coli cells is described. The procedure allows for robust production of highly pure PARP-1 that is free of DNA contamin ...

This chapter describes some of the techniques in use in our laboratories for the investigation of PARP inhibitors in clinical medicine. More specifically, we are involved in investigating the utility of PARP inhibitors in the treatment of hematopoietic malignancies. We are also active ...

The purification of Poly(ADP-ribose) polymerases from overexpressing cells (Sf9 insect cells, Escherichia coli) has been updated to a fast and reproducible three chromatographic steps protocol. After cell lysis, proteins from the crude extract are separated on a Heparine Sepharo ...

Poly(ADP-ribose) polymerases (PARP) participate in diverse biological processes contributing to cellular homeostasis or exacerbating injury. PARP catalyzes the addition of ADP-ribose molecules (pADPr) to the target proteins, a process termed poly-ADP-ribosylation. Ov ...

Poly(ADP-ribose) polymerases have been linked to several cellular functions, most of which being mediated through the dynamics of poly(ADP-ribose) (pADPr). In several pathways, pADPr is the effector molecule that regulates cellular signaling and dictates biological outcomes. p ...

PARP1 can modify a variety of proteins through conserved domains in noncovalent manner. Since poly(ADP-ribose) is highly negatively charged and has a strong binding affinity for its target proteins, noncovalent binding by poly(ADP-ribose) modulates the protein activity during dev ...

Compared to most common posttranslational modifications of proteins, a peculiarity of poly(ADP-ribosyl)ation is the molecular heterogeneity and complexity of the reaction product, poly(ADP-ribose) (PAR). In fact, protein-bound PAR consists of variously sized (2–200 ADP-rib ...

ADP-ribosylation is a well-known post-translational protein modification, which regulates a variety of �cellular processes. The proteins able to catalyze mono- or poly ADP-ribosylation of proteins belong to the family of ADP-ribosyltransferases. A variety of nuclear proteins ...

The enzymatic function of poly(adenosine diphosphate (ADP)-ribose) polymerase (PARP) is central to many of its function as a component of DNA repair machinery, modulator of gene transcription, and cell differentiation. While the auto-modification domain of PARP has been shown to be a pri ...

The structural integrity of eukaryotic genomes, to a great extent, depends on highly regulated and �coordinated enzymatic chromosomal poly(ADP-ribosyl)ation cycles that target chromatin proteins for specific covalent epigenetic poly(ADP-ribose) modification. As a resu ...

An accurate and sensitive detection of catalytic activation of poly(ADP-ribose) polymerase-1 (PARP-1) is required to be performed in a wide variety of samples because this activity plays a role in various cellular responses to DNA damage ranging from DNA repair to cell death, as well as in housek ...

PARP-1 is the most abundantly expressed member of a family of proteins that catalyze the transfer of ADP-ribose units from NAD+ to target proteins. Numerous studies on PARP-1 have revealed its critical roles in regulating various molecular and cellular processes including DNA damage dete ...

Poly (ADP-ribosyl)ation of proteins is a posttranslational modification mediated by poly (ADP-ribose) polymerases (PARPs) that uses NAD+ as substrate to form the negatively charged polymer of poly (ADP-ribose) (PAR). After DNA damage, PARP-1 is responsible for approximately 90% of the ...

The breast cancer 1 and 2, early onset (BRCA1 and BRCA2) genes are important for double-strand break repair by homologous recombination. Cells with inactivating mutations of the BRCA1 or BRCA2 tumor suppressor genes show increased sensitivity to Poly-ADP ribose polymerase (PARP)-inh ...

Generation of the mutated versions of proteins deficient for poly(ADP-ribosyl)ation (PARylation) is a major challenge because there is no clearly defined consensus site for PARylation. In this chapter, we describe possible strategies to produce such mutants, demonstrated by the ex ...

Poly(ADP-ribosyl)ation is a post-translational modification of proteins mediated by Poly(ADP-ribose) polymerases (Parps), a family of 17 members. Among them, Poly(ADP-ribose) polymerase-1 (Parp-1) and Parp-2 are so far the sole enzymes whose catalytic activity has been shown to be ind ...

Studies suggest that inhibiting the hydrolysis of poly(ADP-ribose) (PAR) by targeting the enzyme PAR glycohydrolase (PARG) is a potential chemotherapeutic strategy to induce cell death. However, the lack of structural data for PARG has hindered the discovery of specific PARG inhibit ...

PolyADP-ribosylation plays an essential function in maintenance of genomic stability and cell survival. Although there are several proteins served as acceptor proteins in vitro, there are few proteins in vivo that are identified, including poly(ADP-ribose) polymerase-1. We have b ...

The aim of this chapter is to present feasible strategies for producing novel Parp knock out mice as well multiple knock outs utilizing genetrap and specifically gene targeted ES cell clones publically available through international programs such as KOMP and IGTC. Specifically, we first ...

Poly ADP-ribosylation (PARylation) is a posttranslational protein modification catalyzed by poly �ADP-ribose polymerases (PARPs). Poly ADP-ribose metabolism is involved in a wide range of biological processes, such as maintenance of genome stability, transcriptional reg ...