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Mol Biol (Mosk) ; 48(1): 176-84, 2014.
Article in Russian | MEDLINE | ID: mdl-25842838

ABSTRACT

The Ocr protein, encoded by 0.3 (ocr) gene of bacteriophage T7, belongs to the family of antirestriction proteins that specifically inhibit the type I restriction-modification systems. Native Ocr forms homodimer (Ocr)2 both in solution and in the crystalline state. The Ocr protein belongs to the family of mimicry proteins. F53D A57E and E53R V77D mutant proteins were obtained, which form monomers. It was shown that the values of the dissociation constants Kd for Ocr, Ocr F53D A57E and Ocr F53RV77D proteins with EcoKI enzyme differ in 1000 times: Kd (Ocr) = 10(-10) M, Kd (Ocr F53D A57E and Ocr F53R V77D) = 10(-7) M. Antimodification activity of the Ocr monomeric forms is significantly reduced. We have shown, that Ocr dimeric form has fundamental importance for high inhibitory activity.


Subject(s)
Bacteriophage T7/metabolism , DNA Restriction Enzymes/antagonists & inhibitors , DNA Restriction-Modification Enzymes/metabolism , Escherichia coli K12/metabolism , Protein Multimerization , Viral Proteins/metabolism , Amino Acid Substitution , Bacteriophage T7/genetics , DNA Restriction Enzymes/genetics , DNA Restriction Enzymes/metabolism , DNA Restriction-Modification Enzymes/genetics , Escherichia coli K12/genetics , Escherichia coli K12/virology , Mutation, Missense , Viral Proteins/genetics
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