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[Modification of the modification-restriction system in staphylococci].

A new system of host specificity of DNA, called Sau67 according to the available nomenclature, was identified in Staphylococcus aureus 6782 strain by means of cross titration with staphylophage 729 considering that the phage exhibited the highly effective absorption properties. A total preparation of Sau67 methylases was isolated using ammonium sulfate fractionation. The enzyme preparation contained methylases of cytosine and adenine, where the activity of adenine methylases constituted only for 5% of the total methylase activity. As shown by kinetics of methylation low content of unspecific cellular nucleases was found in the St. aureus 6782 strain; these reasons are important for isolation of restricting endonucleases containing in the strain. 100 micrograms of protein of the total enzymatic fraction enabled to methylate the acceptory DNA at the maximal rate within 1.5 hr of incubation in phosphate buffer, pH 7.9. The fraction of cytosine methylases free of adenine methylating activity was obtained after chromatography on Sepharose blue with NaCl concentration stepwise gradient.

Chromatography, Agarose↗

[Influence of the structure of photoreactive ATP analogs on the affinity modification of phenylalanyl-tRNA synsthetase. Modification of the enzyme at two types of nucleotide sites].

ATP gamma-(p-azidoanilidate) (1) and ATP gamma-(p-azidobenzyl)-methylanilidate (2) were shown to be competitive inhibitors for ATP and amino acid in tRNA aminoacylation catalyzed by E. coli MRE-600 phenylalanyl-tRNA synthetase (E.C.6.1.1.20). Low concentration (10(-5)--10(-6) M) of either ATP, gamma-anilidate or GMP stimulates the aminoacylation of tRNA suggesting their interaction with some nucleotide binding sites of the enzyme other than catalytic ones. Covalent photobinding of (1) to the enzyme does not inhibit aminoacylation, nor does it prevent nucleotides from activating the enzyme. UV-irradiation of the synthetase in the presence of (2) results in complete inactivation of the enzyme which can be prevented by phenylalanine or phenylalanine-ATP to save 50% of the enzyme activity but not ATP and tRNA. The photobinding of (2) to the enzyme in the presence of phenylalanine and ATP removes the activation of the enzyme by nucleotides suggesting that both the catalytic and effector sites of the synthetase are blocked in the same manner by compound (2).

Adenosine Triphosphate↗