Mechanistic dissection of increased enzymatic rate in a phase-separated compartment.
Mechanistic dissection of increased enzymatic rate in a phase-separated compartment.
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在相分离的隔间中对酶速率增加的机械剖析。
DOI：
10.1038/s41589-021-00801-x
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发表时间：
2021-06
影响因子：
14.8
通讯作者：
Rosen MK
中科院分区：
文献类型：
--
作者：
Peeples W;Rosen MK
生物分子浓缩物将大分子浓缩成离散的细胞灶，而没有包封膜。缩合物通常被认为通过增加酶和底物的浓度来增加酶促反应速率（质量作用），尽管这种想法尚未被广泛测试，并且其他调节机制是可能的。在这里，我们描述了一个合成系统，其中SUMO化酶级联被招募到工程冷凝物产生的液-液相分离的多结构域支架蛋白。SUMO化率可以增加高达36倍，在这些液滴相比，周围的散装，这取决于基板KM。这种依赖性在不同底物之间产生了实质性的特异性。高于和低于相分离阈值的反应的分析导致一个定量模型，在该模型中，反应在冷凝物中被加速的质量作用和基板KM的变化，可能是由于支架诱导的分子组织。因此，缩合物可以通过浓缩分子和物理组织它们来调节反应速率。
Biomolecular condensates concentrate macromolecules into discrete cellular foci without an encapsulating membrane. Condensates are often presumed to increase enzymatic reaction rates through increased concentrations of enzymes and substrates (mass action), although this idea has not been widely tested and other mechanisms of modulation are possible. Here we describe a synthetic system where the SUMOylation enzyme cascade is recruited into engineered condensates generated by liquid-liquid phase separation of multidomain scaffolding proteins. SUMOylation rates can be increased up to 36-fold in these droplets compared to the surrounding bulk, depending on substrate KM. This dependency produces substantial specificity among different substrates. Analyses of reactions above and below the phase separation threshold lead to a quantitative model in which reactions in condensates are accelerated by mass action and by changes in substrate KM, likely due to scaffold-induced molecular organization. Thus, condensates can modulate reaction rates both by concentrating molecules and by physically organizing them.
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影响因子：
3.2
作者：
Gao, Yingning;Roberts, Christopher C.;Wheeldon, Ian
通讯作者：
Wheeldon, Ian
影响因子：
9.5
作者：
Kojima T;Takayama S
通讯作者：
Takayama S
DOI：
10.1073/pnas.1706197114
发表时间：
2017-09-26
影响因子：
11.1
作者：
Brady, Jacob P.;Farber, Patrick J.;Kay, Lewis E.
通讯作者：
Kay, Lewis E.
影响因子：
46.9
作者：
Castellana M;Wilson MZ;Xu Y;Joshi P;Cristea IM;Rabinowitz JD;Gitai Z;Wingreen NS
通讯作者：
Wingreen NS
影响因子：
16.6
作者：
Drobot B;Iglesias-Artola JM;Le Vay K;Mayr V;Kar M;Kreysing M;Mutschler H;Tang TD
通讯作者：
Tang TD