山东大学生物化学精品课程 082 The enzyMology of DNA repliCATion.pdfVIP

生物化学 Chapter 8 DNA的生物合成 Chapter 8 DNA的生物合成 Biosythesis of DNA Biosythesis of DNA 1.5 DNA的复制酶体系及其作用机理 The Enzymology of DNA Replication DNA polymerase I (from E.coli cells, consisting a single polypeptide chain) was the first DNA polymerase purified and characterized (by Arthur Kornberg in 1955) 1.5.1 DNA聚合酶 (以DNA为模板的DNA聚合酶) 模板链结合部位 识别并结合模板 引物链结合部位 识别并结合引物 酶活性部位 的复杂性 dNTP专一性结合部位 识别并结合4种dNTP 多种酶活性催化部位 三种酶活性 1.5.1 DNA聚合酶 DNA polymerase I 同时具有多种活性： 1.The polymerase is able to add dNTP to the 3’-hydroxyl group of a preexisting primer strand ( 5’→3’ polymerizing activity); • DNA polymerase I catalyzes the nucleophilic attack by the 3’-OH 1.5.1 DNA聚合酶 3’5’外切酶位点 P414 1.5.1 DNA聚合酶 DNA polymerase I 同时具有多种活性： 2. DNA polymerase I has a separate intrinsic 3’→5’ exonuclease activity, found to be used for correcting mistakes (proofreading) in newly synthesized DNA chains; 1.5.1 DNA聚合酶 3’→5’ exonuclease activity Proof reading activity is slow compared to polymerizing activity, but the stalling of DNAP I after insertion of an incorrect base allows the proofreading activity to catch up with the polymerizing activity and remove the incorrect base. 1.5.1 DNA聚合酶 DNA polymerase I 同时具有多种活性： 1. The polymerase is able to add dNTP to the 3’-h