Temperature-dependent hydrogen deuterium exchange shows impact of analog binding on adenosine deaminase flexibility but not embedded thermal networks.
Temperature-dependent hydrogen deuterium exchange shows impact of analog binding on adenosine deaminase flexibility but not embedded thermal networks.
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DOI：
10.1016/j.jbc.2022.102350
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发表时间：
2022-09
影响因子：
4.8
通讯作者：
Klinman, Judith P.
中科院分区：
文献类型：
--
作者：
Gao, Shuaihua;Zhang, Wenju;Barrow, Samuel L.;Iavarone, Anthony T.;Klinman, Judith P.
通过质谱法分析氢氘交换与温度和突变的关系，已成为蛋白质网络空间分辨率的通用且有效的工具，这些蛋白质网络被认为在催化的热激活中发挥作用。在这项工作中，我们扩展温度依赖性氢氘交换从脱辅基酶结构蛋白质配体复合物。使用腺苷脱氨酶作为原型，我们比较了底物类似物（1-deaza-adenosine）和非常紧密结合的抑制剂/过渡态类似物（喷司他丁）在单一和多个温度下的影响。在一个单一的温度下，我们观察到不同的氢氘交换质谱特性的两个配体，从他们的结合强度的106倍的差异预期。相比之下，温度依赖性氢氘交换质谱的类似模式出现在1-脱氮-腺苷和喷司他丁的存在下，表明任何配体对局部蛋白质展开的生物屏障的类似影响。我们在喷司他丁存在下将温度依赖性氢氘交换扩展到腺苷脱氨酶的功能改变突变体，并揭示了与先前报道的脱辅基酶高度相似的蛋白质热网络（Gao等人，2020，JACS 142，19936-19949）。最后，我们讨论了喷司他丁结合整体蛋白质的灵活性与特定位点的热传递途径的背景下，底物诱导的变化的分布式蛋白质构象景观，协同作用与嵌入式蛋白质热网络，以实现有效的催化模型的差异影响。
The analysis of hydrogen deuterium exchange by mass spectrometry as a function of temperature and mutation has emerged as a generic and efficient tool for the spatial resolution of protein networks that are proposed to function in the thermal activation of catalysis. In this work, we extend temperature-dependent hydrogen deuterium exchange from apo-enzyme structures to protein–ligand complexes. Using adenosine deaminase as a prototype, we compared the impacts of a substrate analog (1-deaza-adenosine) and a very tight-binding inhibitor/transition state analog (pentostatin) at single and multiple temperatures. At a single temperature, we observed different hydrogen deuterium exchange-mass spectrometry properties for the two ligands, as expected from their 106-fold differences in strength of binding. By contrast, analogous patterns for temperature-dependent hydrogen deuterium exchange mass spectrometry emerge in the presence of both 1-deaza-adenosine and pentostatin, indicating similar impacts of either ligand on the enthalpic barriers for local protein unfolding. We extended temperature-dependent hydrogen deuterium exchange to a function-altering mutant of adenosine deaminase in the presence of pentostatin and revealed a protein thermal network that is highly similar to that previously reported for the apo-enzyme (Gao et al., 2020, JACS 142, 19936-19949). Finally, we discuss the differential impacts of pentostatin binding on overall protein flexibility versus site-specific thermal transfer pathways in the context of models for substrate-induced changes to a distributed protein conformational landscape that act in synergy with embedded protein thermal networks to achieve efficient catalysis.
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影响因子：
16.6
作者：
Fried SD;Boxer SG
通讯作者：
Boxer SG
DOI：
10.1039/c8cc02426j
发表时间：
2018-06-19
期刊：
Chemical communications (Cambridge, England)
影响因子：
--
作者：
Maria-Solano MA ;Serrano-Hervás E ;Romero-Rivera A ;Iglesias-Fernández J ;Osuna S
通讯作者：
Osuna S
通过氢交换质谱法对多蛋白信号组件的构象见解。
影响因子：
6.8
作者：
Harrison, Rane A.;Engen, John R.
通讯作者：
Engen, John R.
DOI：
10.1016/j.jasms.2006.06.006
发表时间：
2006-11-01
影响因子：
3.2
作者：
Englander, S. Walter
通讯作者：
Englander, S. Walter
影响因子：
48
作者：
Masson, Glenn R.;Burke, John E.;Rand, Kasper D.
通讯作者：
Rand, Kasper D.