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Interaction of Endonuclease Eco RI with Short Specific and Nonspecific Oligonucleotides
Author(s) -
Kolocheva Tat'yana I.,
Maksakova Galiya A.,
Bugreev Dmitry D.,
Nevinsky Georgy A.
Publication year - 2001
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/152165401753544269
Subject(s) - ecori , chemistry , endonuclease , deoxyribose , stereochemistry , oligonucleotide , nucleotide , ligand (biochemistry) , enzyme , nucleic acid , dna , biochemistry , restriction enzyme , receptor , gene
The interaction of Eco RI with different oligodeoxyribonucleotides (ODNs) was analyzed using the method of the slow step‐by‐step simplification in their complexity. Orthophosphate (K I = 31 mM), 2‐deoxyribose 5‐phosphate (K I = 4.6 mM) and different dNMPs (K I = 2.1‐2.5 mM) were shown to be the minimal 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 2.0. Weak nonspecific electrostatic contacts of Eco RI with internucleotide phosphate groups of ODNs can account for about 5 orders of magnitude in the ligand affinity, whereas the contribution of specific interactions between Eco RI and d(pN) n is no more than 2 orders of magnitude of a total ODN's affinity.