TAILIEUCHUNG - Báo cáo khoa học: Catalytic transformations of supercoiled DNA as studied by flow linear dichroism technique

A catalytic turnover of supercoiled DNA (scDNA) transformation medi-ated by topoisomerases leads to changes in the linking number (Lk) of the polymeric substrate by 1 or 2 per cycle. As a substrate of the topoisomeri-zation reaction it is chemically identical to its product; even a single catalytic event results in the quantum leap in the scDNA topology. | iFEBS Journal Catalytic transformations of supercoiled DNA as studied by flow linear dichroism technique Alexander Gabibov1 2 3 Elena Yakubovskaya1 Mark Lukin4 Peter Favorov3 Andrey Reshetnyak5 and Michael Monastyrsky2 6 1 Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences Moscow Russia 2 Max-Plank-Institute for Physics of Complex Systems Dresden Germany 3 Engelhardt Institute of Molecular Biology Russian Academy of Sciences Moscow Russia 4 Institute of ExperimentalCardiology Cardiology Research Center Moscow Russia 5 Moscow State University ChemicalDepartment Vorobjovy Gory Moscow Russia 6 Institute of Theoreticaland ExperimentalPhysics Russian Academy of Sciences Moscow Russia Keywords anticancer drugs flow linear dichroism supercoiled DNA transformations topoisomerases Correspondence A. Gabibov Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry Russian Academy of Sciences 16 10 Mikluho-Maklaya str. 117871 Moscow Russia Fax 7095 3107007 Tel 7095 4298269 E-mail gabibov@ Note A. G. and M. M. were visiting scientists. Received 28 February 2005 revised 13 September 2005 accepted 19 October 2005 A catalytic turnover of supercoiled DNA scDNA transformation mediated by topoisomerases leads to changes in the linking number Lk of the polymeric substrate by 1 or 2 per cycle. As a substrate of the topoisomerization reaction it is chemically identical to its product even a single catalytic event results in the quantum leap in the scDNA topology. Non-intrusive continuous assay to measure the kinetics of the scDNA topoisomerization was performed. The development of such a technique was hindered because of multiple DNA species of intermediate topology present in the reaction mixture. The interrelation of DNA topology its hydrodynamics and optical anisotropy enable us to use the flow linear dichroism technique FLD for continuous monitoring of the scDNA topoisomerization reaction. This approach permits us to study the kinetics

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