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Cross-linking of SsoII restriction endonuclease to cognate and non-cognate DNAs
Sheflyan GYa1, E A Kubareva, S A Kuznetsova
1A.N. Belozersky Institute of Physical and Chemical Biology, Moscow State University, Russian Federation.
Abstract:
Specific and non-specific interactions of SsoII restriction endonuclease (R.SsoII) were probed by the method of covalent attachment to modified DNA containing an active monosubstituted pyrophosphate internucleotide bond instead of a phosphodiester one. R.SsoII with six N-terminal His residues was shown to be cross-linked to duplexes with this type of modification, either containing or not the recognition sequence. Competition experiments with covalent attachment of R.SsoII to activated DNAs demonstrated the similar affinity of the enzyme to cognate and non-cognate DNAs in the absence of cofactor, Mg2+ ions.