Coenzyme binding in F420-dependent secondary alcohol dehydrogenase, a member of the bacterial luciferase family
Coenzyme binding in F420-dependent secondary alcohol dehydrogenase, a member of the bacterial luciferase family
复制标题
DOI：
10.1016/j.str.2004.02.010
复制
发表时间：
2004-03-01
期刊：
影响因子：
5.7
通讯作者：
Ermler, U
中科院分区：
文献类型：
--
作者：
Aufhammer, SW;Warkentin, E;Ermler, U
来自产甲烷古菌的 F-420 依赖性仲醇脱氢酶 (Adf) 是正在生长的细菌荧光素酶家族的成员，该家族都是 TIM 桶酶，其中大多数具有不寻常的非脯氨酰顺式肽键。我们在此报告了来自嗜热甲烷库菌的 Adf 与 F-420-丙酮加合物复合物的晶体结构，分辨率为 1.8 埃。 Adf 中 F-420 结合模式的知识为将 F-420 和 FMN 建模到该家族的其他酶中提供了分子基础。非脯氨酰顺式肽键被确定为凸起的重要部分，该凸起充当 F-420 Re-face 的支撑，以使其保持弯曲构象。 F-420-丙酮加合物的丙酮部分位于深埋在蛋白质内部的 F-420 的 Si 面。可以可靠地对异丙醇进行建模并假设氢转移机制。 His39 和 Glu108 可以被确定为丙酮或异丙醇氧结合和催化的关键参与者。
F-420-dependent secondary alcohol dehydrogenase (Adf) from methanogenic archaea is a member of the growing bacterial luciferase family which are all TIM barrel enzymes, most of which with an unusual non-prolyl cis peptide bond. We report here on the crystal structure of Adf from Methanoculleus thermophilicus at 1.8 Angstrom resolution in complex with a F-420-acetone adduct. The knowledge of the F-420 binding mode in Adf provides the molecular basis for modeling F-420 and FMN into the other enzymes of the family. A non-prolyl cis peptide bond was identified as an essential part of a bulge that serves as backstop at the Re-face of F-420 to keep it in a bent conformation. The acetone moiety of the F-420-acetone adduct is positioned at the Si-face of F-420 deeply buried inside the protein. Isopropanol can be reliably modeled and a hydrogen transfer mechanism postulated. His39 and Glu108 can be identified as key players for binding of the acetone or isopropanol oxygens and for catalysis.