Saha, Swati and Rao, Desirazu N (1995) ATP hydrolysis is required for DNA cleavage by EcoPI restriction enzyme. In: Journal of Molecular Biology, 247 (4). pp. 559-567.
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The type III restriction endonuclease EcoPI, coded by bacteriophage Fl, cleaves unmodified DNA in the presence of ATP and magnesium ions. We show that purified EcoPI restriction enzyme fails to cleave DNA in the presence of non-hydrolyzable ATP analogs. More importantly, this study demonstrates that EcoPI restriction enzyme has an inherent ATPase activity, and ATP hydrolysis is necessary for DNA cleavage. Furthermore, we show that the progress curve of the reaction with Eco PI restriction enzyme exhibits a lag which is dependent on the enzyme concentration. Kinetic analysis of the progress curves of the reaction suggest slow transitions that can occur during the reaction, characteristic of hysteretic enzymes. The role of ATP in the cleavage mechanism of type III restriction enzymes is discussed.
|Item Type:||Journal Article|
|Additional Information:||Copyright of this article belongs to Elsevier Science.|
|Keywords:||type III restriction-modification;ATP hydrolysis;DNA cleavage;hysteresis|
|Department/Centre:||Division of Biological Sciences > Biochemistry|
|Date Deposited:||26 Apr 2011 09:51|
|Last Modified:||26 Apr 2011 09:51|
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