Dynamic light scattering study of base excision DNA repair proteins and their complexes
| Autor: | Inna A. Vasil'eva, Olga I. Lavrik, N. A. Moor, Rashid O. Anarbaev |
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| Rok vydání: | 2019 |
| Předmět: |
DNA Repair
DNA polymerase Poly (ADP-Ribose) Polymerase-1 Biophysics Biochemistry Analytical Chemistry Protein–protein interaction 03 medical and health sciences XRCC1 Dynamic light scattering DNA-(Apurinic or Apyrimidinic Site) Lyase AP site Molecular Biology DNA Polymerase beta Polymerase 030304 developmental biology 0303 health sciences biology Phosphoric Diester Hydrolases Chemistry 030302 biochemistry & molecular biology Base excision repair Heterotetramer Dynamic Light Scattering X-ray Repair Cross Complementing Protein 1 biology.protein |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1867:297-305 |
| ISSN: | 1570-9639 |
| DOI: | 10.1016/j.bbapap.2018.10.009 |
| Popis: | Base excision repair (BER) involves many enzymes acting in a coordinated fashion at the most common types of DNA damage. The coordination is facilitated by interactions between the enzymes and accessory proteins, X-ray repair cross-complementing protein 1 (XRCC1) and poly(ADP-ribose) polymerase 1 (PARP1). Here we use dynamic light scattering (DLS) technique to determine the hydrodynamic sizes of several BER enzymes and proteins, DNA polymerase β (Polβ), apurinic/apyrimidinic endonuclease 1 (APE1), tyrosyl-DNA phosphodiesterase 1 (TDP1), XRCC1 and PARP1, present alone or in the equimolar mixtures with each other. From the DLS data combined with glutaraldehyde cross-linking experiments and previous quantitative binding data the oligomeric states of BER proteins and their complexes are estimated. All the proteins have been proposed to form homodimers upon their self-association. The most probable oligomerization state of the binary complexes formed by PARP1 with various proteins is a heterotetramer. The oligomerization state of the binary complexes formed by XRCC1 varies from heterodimer to heterotetramer, depending on the partner. The DLS technique is applied for the first time to measure the hydrodynamic sizes of PARP1 molecules covalently bound with poly(ADP-ribose) (PAR) synthesized upon the automodification reaction. PARP1 has been detected to form huge conglomerates stabilized by Mg2+ coordinated bonds with PAR polymers. |
| Databáze: | OpenAIRE |
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