Müller Joachim, Niemeyer Christof M
Fakultät Chemie, Technische Universität Dortmund, Biologisch-Chemische Mikrostrukturtechnik, Dortmund, Germany.
Biochem Biophys Res Commun. 2008 Dec 5;377(1):62-7. doi: 10.1016/j.bbrc.2008.09.078. Epub 2008 Sep 25.
This study aims to establish model systems for the exploration of proximity effects, occurring in natural multienzyme complexes. DNA-directed assembly of covalent conjugates of DNA oligonucleotides and Glucose Oxidase (GOX) or Horseradish peroxidase (HRP) was used to generate supramolecular complexes, in which the two enzymes were arranged with defined spatial orientation. Electrophoretic studies indicated that the assembly efficiency significantly depends on positional and sterical factors of the two DNA-enzyme conjugates. Kinetic rate measurements of the coupled reaction of glucose oxidation and Amplex Red peroxidation were carried out with microplate-immobilized DNA-GOX-HRP complexes, and the influence of Catalase on this reaction was determined. The kinetic measurements revealed a significant increase in the reactivity of the complexes, in which GOX and HRP were immobilized in direct proximity on a complementary DNA carrier.
本研究旨在建立用于探索天然多酶复合物中发生的邻近效应的模型系统。利用DNA寡核苷酸与葡萄糖氧化酶（GOX）或辣根过氧化物酶（HRP）的共价缀合物的DNA定向组装来生成超分子复合物，其中两种酶以确定的空间取向排列。电泳研究表明，组装效率显著取决于两种DNA-酶缀合物的位置和空间因素。用微孔板固定的DNA-GOX-HRP复合物进行了葡萄糖氧化和Amplex Red过氧化偶联反应的动力学速率测量，并确定了过氧化氢酶对该反应的影响。动力学测量结果显示，当GOX和HRP直接邻近固定在互补DNA载体上时，复合物的反应活性显著增加。
