Interaction of endonuclease EcoRI with short specific and nonspecific oligonucleotides

Citation
Ti. Kolocheva et al., Interaction of endonuclease EcoRI with short specific and nonspecific oligonucleotides, IUBMB LIFE, 51(3), 2001, pp. 189-195
Citations number
31
Categorie Soggetti
Biochemistry & Biophysics
Journal title
IUBMB LIFE
ISSN journal
15216543 → ACNP
Volume
51
Issue
3
Year of publication
2001
Pages
189 - 195
Database
ISI
SICI code
1521-6543(200103)51:3<189:IOEEWS>2.0.ZU;2-A
Abstract
The interaction of EcoRI with different oligodeoxyribonucleotides (ODNs) wa s analyzed using the method of the slow step-by-step simplification in thei r complexity. Orthophosphate (K-I = 31 mM), 2-deoxyribose 5-phosphate (K-I = 4.6 mM) and different dN-MPs (K-I = 2.1-2.5 mM) were shown to be the mini mal ligands of the enzyme. The lengthening of a nonspecific d(pN)(n) (n = 1 -6) by one nucleotide unit resulted in the increase of their affinity by a factor of similar to2.0. Weak nonspecific electrostatic contacts of EcoRI w ith internucleotide phosphate groups of ODNs can account for about 5 orders of magnitude in the ligand affinity, whereas the contribution of specific interactions between EcoRI and d(pN), is no more than 2 orders of magnitude of a total ODN's affinity.