Structures of Mycobacterium tuberculosis isoprenyl diphosphate synthase Rv2173 in substrate-bound forms.

IF 1.1 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS
James A Titterington, Ngoc Anh Thu Ho, Charles P H Beasley, Francis Mann, Edward N Baker, Timothy M Allison, Jodie M Johnston
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引用次数: 0

Abstract

We report structures of the Mycobacterium tuberculosis isoprenyl diphosphate synthase Rv2173 in three forms: apo and two substrate-bound forms [isoprenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP)]. The protein possesses a canonical all-α-helical trans-isoprenyl diphosphate synthase fold that is dimeric in each form. There are some differences between the structures: the IPP-bound form shows IPP bound in the DMAPP/allylic substrate-binding site with three divalent metal ions bound around the IPP and the complete C-terminus closing around the active site, while the apo and DMAPP-bound forms are more open, with some of the C-terminal region disordered, supporting suggestions that the C-terminus is important in substrate entry/product exit. In the DMAPP form DMAPP occupies the expected allylic substrate site, but only two metal ions are associated with the binding, with the DMAPP diphosphates adopting a slightly different binding pose compared with IPP in the same site, and the third metal-binding site is unoccupied. In no case is the IPP binding site occupied by IPP. There has been some uncertainty regarding product length for Rv2173, with variable lengths being reported. In the structures reported here, the `capping' residue at the bottom of the binding cavity is tryptophan and comparison with other IPP synthases suggests that the structure of Rv2173 is most consistent with a C10-C15 final product size.

底物结合形式的结核分枝杆菌异戊二酯二磷酸合酶Rv2173的结构。
我们报告了结核分枝杆菌异戊烯基二磷酸合酶 Rv2173 的三种结构形式:apo 和两种底物结合形式 [异戊烯基二磷酸(IPP)和二甲基烯丙基二磷酸(DMAPP)]。该蛋白具有典型的全α-螺旋反式异戊烯基二磷酸合成酶折叠,每种形式都是二聚体。这两种形式的结构存在一些差异:与 IPP 结合的形式显示 IPP 与 DMAPP/烯丙基底物结合位点结合,三个二价金属离子结合在 IPP 周围,整个 C 端封闭在活性位点周围;而与 apo 和 DMAPP 结合的形式则更为开放,部分 C 端区域紊乱,这支持了 C 端在底物进入/产物排出中起重要作用的观点。在 DMAPP 形式中,DMAPP 占据了预期的烯丙基底物位点,但只有两个金属离子与之结合,与 IPP 相比,DMAPP 二磷酸盐在同一位点的结合姿态略有不同,第三个金属结合位点未被占据。在任何情况下,IPP 结合位点都不会被 IPP 占用。关于 Rv2173 的产物长度一直存在一些不确定性,有报告称其长度不一。在本文报告的结构中,结合空腔底部的 "盖帽 "残基是色氨酸,与其他 IPP 合成酶的比较表明,Rv2173 的结构最符合 C10-C15 的最终产物大小。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Acta crystallographica. Section F, Structural biology communications
Acta crystallographica. Section F, Structural biology communications BIOCHEMICAL RESEARCH METHODSBIOCHEMISTRY &-BIOCHEMISTRY & MOLECULAR BIOLOGY
CiteScore
1.90
自引率
0.00%
发文量
95
期刊介绍: Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.
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