Logo Medical Science Monitor

Call: +1.631.470.9640
Mon - Fri 10:00 am - 02:00 pm EST

Contact Us

Logo Medical Science Monitor Logo Medical Science Monitor Logo Medical Science Monitor

02 May 2003

Studies of PARP-1: a chemical biology approach

P. Gao, H. Liu

Med Sci Monit 2003; 9(1): 26-0 :: ID: 15077

Abstract

A set of gene constructed encoding the wild-type PARP, the various combinations of the individual domains of PARP, and several mutations of the individual domains have been obtained. All these genes are overexpressed in E. coli, and the purified proteins are used in different combinations to assess their competence as catalyst or as acceptors in poly(ADP-ribosyl)ation reactions. One truncated PARP mutant showed no self-modification activity and no catalytic activities when histone or PARP automodification domain are used as substrates. However, this mutant is able to use all PARP fragments that contain intact DNA-binding domain as substrates for poly(ADP-ribosyl)ation reactions. These results not only reveal the importance of molecular recognition between PARP and its targets, but also indicate the existence of additional modification sites at the C-terminal of PARP DNA binding domain.To gain further insight into the modification sites on PARP and its substrate proteins, and to learn more about the properties of the ADP-ribose polymer, we also synthesized several fluorinated NAD analogues and applied them to poly(ADP-ribosyl)ation reactions. One of these analogues, 2’-deoxy-2’-F-NAD+ in which the 2’ hydroxyl of nicotinamide ribose of NAD+ is substituted by a fluoro atom, should inhibit the branching reaction of PARP. As expected, our results indicate that with this analogue as co-substrate, PARP modifies the substrate proteins by forming non-branched polymers which can be recognized by anti-PAR antibody. This analogue provides us an easy way to distinguish types of modified proteins and will be useful in searching for PARP substrates. These studies and results obtained from our ongoing research with the NAD+ analogues in biological applications will be presented. References: 1.Griesenbeck J, Oei SL, Mayer-Kuckuk P, Ziegler M, Buchlow G, Schweiger M: Biochemistry, 1997; 36: 7297 2.de Murcia G, Shall S: From DNA damage and stress signaling to cell death polyADP-ribosylation reactions. Oxford University Press, New York 2000, 35-79 3.Ruf A, Rolli V, de Murcia G, Schulz GE: J Mol Biol, 1998; 278: 57-65

Keywords: truncated PARP, fluorinated NAD+ analogues

Add Comment 0 Comments

Editorial

01 March 2024 : Editorial  

Editorial: First Regulatory Approvals for CRISPR-Cas9 Therapeutic Gene Editing for Sickle Cell Disease and Transfusion-Dependent β-Thalassemia

Dinah V. Parums

DOI: 10.12659/MSM.944204

Med Sci Monit 2024; 30:e944204

0:00

In Press

18 Mar 2024 : Clinical Research  

Sexual Dysfunction in Women After Tibial Fracture: A Retrospective Comparative Study

Med Sci Monit In Press; DOI: 10.12659/MSM.944136  

0:00

21 Feb 2024 : Clinical Research  

Potential Value of HSP90α in Prognosis of Triple-Negative Breast Cancer

Med Sci Monit In Press; DOI: 10.12659/MSM.943049  

22 Feb 2024 : Review article  

Differentiation of Native Vertebral Osteomyelitis: A Comprehensive Review of Imaging Techniques and Future ...

Med Sci Monit In Press; DOI: 10.12659/MSM.943168  

23 Feb 2024 : Clinical Research  

A Study of 60 Patients with Low Back Pain to Compare Outcomes Following Magnetotherapy, Ultrasound, Laser, ...

Med Sci Monit In Press; DOI: 10.12659/MSM.943732  

Most Viewed Current Articles

16 May 2023 : Clinical Research  

Electrophysiological Testing for an Auditory Processing Disorder and Reading Performance in 54 School Stude...

DOI :10.12659/MSM.940387

Med Sci Monit 2023; 29:e940387

0:00

17 Jan 2024 : Review article  

Vaccination Guidelines for Pregnant Women: Addressing COVID-19 and the Omicron Variant

DOI :10.12659/MSM.942799

Med Sci Monit 2024; 30:e942799

0:00

14 Dec 2022 : Clinical Research  

Prevalence and Variability of Allergen-Specific Immunoglobulin E in Patients with Elevated Tryptase Levels

DOI :10.12659/MSM.937990

Med Sci Monit 2022; 28:e937990

0:00

01 Jan 2022 : Editorial  

Editorial: Current Status of Oral Antiviral Drug Treatments for SARS-CoV-2 Infection in Non-Hospitalized Pa...

DOI :10.12659/MSM.935952

Med Sci Monit 2022; 28:e935952

0:00

Your Privacy

We use cookies to ensure the functionality of our website, to personalize content and advertising, to provide social media features, and to analyze our traffic. If you allow us to do so, we also inform our social media, advertising and analysis partners about your use of our website, You can decise for yourself which categories you you want to deny or allow. Please note that based on your settings not all functionalities of the site are available. View our privacy policy.

Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750