Structural basis of bidirectional allostery across the heme in a cytochrome P450 enzyme.
Structural basis of bidirectional allostery across the heme in a cytochrome P450 enzyme.
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DOI：
10.1016/j.jbc.2023.104977
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发表时间：
2023-08
影响因子：
4.8
通讯作者：
Estrada, D. Fernando
中科院分区：
文献类型：
--
作者：
Kumar, Amit;Estrada, D. Fernando
细胞色素P450（CYP）是存在于所有生命界中的含血红素的酶，并且共享结构同源的球状蛋白质折叠。CYP利用血红素远端的结构来识别和协调底物，而与氧化还原伴侣蛋白质的必要相互作用在相对的近端表面介导。在目前的研究中，我们研究了细菌酶CYP 121 A1的血红素功能变构，该酶利用非极性远端到远端二聚体界面特异性结合其二环酪氨酸底物。将氟检测的核磁共振（19 F-NMR）光谱与远端表面残基（FG环的S171 C）、B-螺旋的一个残基（N84 C）和两个近端表面残基（T103 C和T333 C）的位点特异性标记结合，并使用巯基反应性氟标记。肾上腺氧还蛋白被用作替代氧化还原蛋白，并被发现促进FG环的闭合排列，类似于单独添加底物。通过诱变两个CYP 121碱性表面残基破坏蛋白质-蛋白质界面，消除了变构效应。此外，近端表面的19 F-NMR光谱表明，配体诱导的变构调节的环境中的C-螺旋，但不是曲折区域的酶。鉴于该酶家族的高度结构同源性，我们将这项工作的结果解释为代表CYP中保守的变构网络。
Cytochromes P450 (CYPs) are heme-containing enzymes that are present in all kingdoms of life and share a structurally homologous, globular protein fold. CYPs utilize structures distal to the heme to recognize and coordinate substrates, while the necessary interactions with redox partner proteins are mediated at the opposite, proximal surface. In the current study, we investigated the functional allostery across the heme for the bacterial enzyme CYP121A1, which utilizes a non-polar distal-to-distal dimer interface for specific binding of its dicyclotyrosine substrate. Fluorine-detected Nuclear Magnetic Resonance (19F-NMR) spectroscopy was combined with site-specific labeling of a distal surface residue (S171C of the FG-loop), one residue of the B-helix (N84C), and two proximal surface residues (T103C and T333C) with a thiol-reactive fluorine label. Adrenodoxin was used as a substitute redox protein and was found to promote a closed arrangement of the FG-loop, similar to the addition of substrate alone. Disruption of the protein–protein interface by mutagenesis of two CYP121 basic surface residues removed the allosteric effect. Moreover, 19F-NMR spectra of the proximal surface indicate that ligand-induced allostery modulates the environment at the C-helix but not the meander region of the enzyme. In light of the high degree of structural homology in this family of enzymes, we interpret the findings from this work to represent a conserved allosteric network in CYPs.
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影响因子：
5.6
作者：
Fischer A;Smieško M
通讯作者：
Smieško M
影响因子：
4.6
作者：
Kumar A;Campomizzi CS;Jay N;Ferguson S;Scheffler EJ;Lioi J;Tu C;Qu J;Simons C;Estrada DF
通讯作者：
Estrada DF
DOI：
10.1016/j.jbc.2021.100794
发表时间：
2021-01
期刊：
The Journal of biological chemistry
影响因子：
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作者：
Brixius-Anderko S;Scott EE
通讯作者：
Scott EE
影响因子：
15
作者：
Baylon, Javier L.;Lenov, Ivan L.;Sligar, Stephen G.;Tajkhorshid, Emad
通讯作者：
Tajkhorshid, Emad
影响因子：
2.9
作者：
Jay N;Duffy SR;Estrada DF
通讯作者：
Estrada DF