Russian Journal of Bioorganic Chemistry

, Volume 38, Issue 4, pp 376–382 | Cite as

Oligomerization of site-specific nicking endonuclease BspD6I at high protein concentrations

  • S. A. Sekerina
  • A. V. Grishin
  • A. Yu. Ryazanova
  • R. I. Artyukh
  • E. A. Rogulin
  • A. K. Yunusova
  • T. S. Oretskaya
  • L. A. Zheleznaya
  • E. A. Kubareva
Article
  • 63 Downloads

Abstract

The ability of site-specific nicking endonuclease BspD6I (Nt.BspD6I) to oligomerize at concentrations > 0.5 μM (>0.035 mg/mL) is studied. Three states of Nt.BspD6I are registered via electrophoretic studies both in the presence and in the absence of DNA. Estimation of their molecular mass allows assigning them as a monomer, a dimer and a trimer. Both dimeric and monomeric Nt.BspD6I are shown to hydrolyze its DNA substrate with the same specificity. Calculation of the electrostatic potential distribution on the Nt.BspD6I globule surface shows that the protein molecule is a dipole. The Nt.BspD6I oligomeric forms are likely to be the result of ionic protein-protein interactions.

Keywords

heterodimeric restriction endonucleases nicking endonuclease oligomerization protein docking 

Abbreviations

PDB

Protein Data Bank

IPTG

isopropyl-β-D-1-thiogalactopyranoside

NE

nicking endonuclease

Mth

theoretical molecular weight

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Copyright information

© Pleiades Publishing, Ltd. 2012

Authors and Affiliations

  • S. A. Sekerina
    • 1
  • A. V. Grishin
    • 1
  • A. Yu. Ryazanova
    • 1
  • R. I. Artyukh
    • 2
  • E. A. Rogulin
    • 2
  • A. K. Yunusova
    • 2
  • T. S. Oretskaya
    • 1
  • L. A. Zheleznaya
    • 2
  • E. A. Kubareva
    • 1
  1. 1.Chemistry Department, Faculty of Bioengineering and Bioinformatics and A. N. Belozersky Institute of Physico-Chemical BiologyMoscow State UniversityMoscowRussia
  2. 2.Institute of Theoretical and Experimental BiophysicsPushchinoRussia

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