National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM124847
米国
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
GM100008
米国
引用
ジャーナル: Nat Commun / 年: 2019 タイトル: Convergent allostery in ribonucleotide reductase. 著者: William C Thomas / F Phil Brooks / Audrey A Burnim / John-Paul Bacik / JoAnne Stubbe / Jason T Kaelber / James Z Chen / Nozomi Ando / 要旨: Ribonucleotide reductases (RNRs) use a conserved radical-based mechanism to catalyze the conversion of ribonucleotides to deoxyribonucleotides. Within the RNR family, class Ib RNRs are notable for ...Ribonucleotide reductases (RNRs) use a conserved radical-based mechanism to catalyze the conversion of ribonucleotides to deoxyribonucleotides. Within the RNR family, class Ib RNRs are notable for being largely restricted to bacteria, including many pathogens, and for lacking an evolutionarily mobile ATP-cone domain that allosterically controls overall activity. In this study, we report the emergence of a distinct and unexpected mechanism of activity regulation in the sole RNR of the model organism Bacillus subtilis. Using a hypothesis-driven structural approach that combines the strengths of small-angle X-ray scattering (SAXS), crystallography, and cryo-electron microscopy (cryo-EM), we describe the reversible interconversion of six unique structures, including a flexible active tetramer and two inhibited helical filaments. These structures reveal the conformational gymnastics necessary for RNR activity and the molecular basis for its control via an evolutionarily convergent form of allostery.
全体 : Inhibited filament of ribonucleoside-diphosphate reductase compos...
全体
名称: Inhibited filament of ribonucleoside-diphosphate reductase composed of NrdE alpha subunit
要素
複合体: Inhibited filament of ribonucleoside-diphosphate reductase composed of NrdE alpha subunit
タンパク質・ペプチド: Ribonucleoside-diphosphate reductase
リガンド: 2'-DEOXYADENOSINE 5'-TRIPHOSPHATE
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超分子 #1: Inhibited filament of ribonucleoside-diphosphate reductase compos...
超分子
名称: Inhibited filament of ribonucleoside-diphosphate reductase composed of NrdE alpha subunit タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1 詳細: The filament is a double helix. Each helix is composed of NrdE subunits dimerizing at alternating canonical and non-canonical interfaces.
凍結剤: ETHANE / チャンバー内湿度: 90 % / チャンバー内温度: 300 K / 装置: FEI VITROBOT MARK IV / 詳細: 3.5 seconds blotting.
詳細
Samples of the dATP-induced NrdE filament were produced by incubating 40 uM holo-NrdE with 100 uM dATP and 1 mM CDP in assay buffer. The mixture was then diluted to 10 uM NrdE in the same nucleotide-containing buffer.