초록

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Abstract ▼ AI-Helper

The closed circular form DNA (Form I) was changed into linear form DNA (form III) by Eco RI endonuclease through the intermediate open-circular form (form II), which could be introduced to the second turn-over, and the intermediate form was more accumulated when the condition was more deviated from ...

The closed circular form DNA (Form I) was changed into linear form DNA (form III) by Eco RI endonuclease through the intermediate open-circular form (form II), which could be introduced to the second turn-over, and the intermediate form was more accumulated when the condition was more deviated from the optimal state. The cleavage of form II and III DNA was more sensitive to inhibition to ethidium bromide (EtBr) than the linearization of form I DNA. The cleavage reactions were stimulated by a low concentration of NaCl, spermidine, and spermine. At high concentration, reactions were markedly inhibited by masking the charge of DNA. Km value of form I DNA was lower than that of form III DNA and thus superhelical form had a higher affinity than linear form DNA. The enzyme activity was sensitive to treatment of functional group blocking agent such as diisopropyl fluorophosphate, iodoacetamide, and diethylpyrocarbonate. The enzyme activity was also decreased in the presence of trinitrobenzene-sulfonate. From these results, it was thought that serine and histidine residues of the enzyme were directly involved in the phosphodiesterase action and the lysine residues might hold the phosphodiester bond of DNA.