The Significance of the Location of Mutations for the Native-State Dynamics of Human Lysozyme.
The Significance of the Location of Mutations for the Native-State Dynamics of Human Lysozyme.
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DOI：
10.1016/j.bpj.2016.10.028
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发表时间：
2016-12-06
影响因子：
3.4
通讯作者：
Kumita, Janet R.
中科院分区：
文献类型：
--
作者：
Ahn, Minkoo;Hagan, Christine L.;Bernardo-Gancedo, Ana;De Genst, Erwin;Newby, Francisco N.;Christodoulou, John;Dhulesia, Anne;Dumoulin, Mireille;Robinson, Carol V.;Dobson, Christopher M.;Kumita, Janet R.
人溶菌酶转化为淀粉样纤维与一种罕见但致命的遗传性非神经性系统性淀粉样变性有关。大量聚集蛋白质的积累被认为是由疾病相关溶菌酶变体的瞬时中间物种的形成引发的，这基本上是由于在生理相关条件下全局协同性的丧失。有趣的是，所有五个天然存在的淀粉样单点突变都位于溶菌酶的β结构域，该区域在瞬时中间物质形成期间主要展开。由于缺乏已知的天然存在的，淀粉样蛋白，单点突变的α-结构域，我们选择了三个特定的突变，以解决的位置可能对天然状态的动力学的影响，通过氢-氘（HD）交换实验研究NMR光谱分析，和质谱。我们比较了不稳定α结构域突变（I23 A）与充分表征的I59 T β结构域变体的影响。我们还研究了在结构域界面（I56 V）对天然状态稳定性影响较小的突变的影响，并将其与C-螺旋（I89 V）内具有类似稳定性的变体进行了比较。我们表明，当变体具有类似的降低的天然状态稳定性时，突变的位置（I23 A与I59 T）对天然状态动态至关重要，其中α结构域突变在生理相关条件下填充瞬时中间物种的能力显着较低。有趣的是，与含有α-结构域突变的变体相比，界面处的突变（I56 V）在促进高温下瞬时中间物质的形成方面具有更大的作用，即使该突变仅导致溶菌酶天然状态稳定性的微小变化。这些研究结果表明，特定突变的位置是一个重要的因素，在确定人类溶菌酶的背景下，其倾向于填充聚集倾向的短暂的中间物种与致病性淀粉样蛋白形成的天然状态的动力学特性。
The conversion of human lysozyme into amyloid fibrils is associated with a rare but fatal hereditary form of nonneuropathic systemic amyloidosis. The accumulation of large amounts of aggregated protein is thought to be initiated by the formation of transient intermediate species of disease-related lysozyme variants, essentially due to the loss of global cooperativity under physiologically relevant conditions. Interestingly, all five naturally occurring, amyloidogenic, single-point mutations are located in the β-domain of lysozyme, the region that is predominantly unfolded during the formation of the transient intermediate species. Given the lack of known naturally occurring, amyloidogenic, single-point mutations in the α-domain, we chose three specific mutations to address the effects that location may have on native-state dynamics, as studied by hydrogen-deuterium (HD) exchange experiments analyzed by NMR spectroscopy, and mass spectrometry. We compared the effect of a destabilizing α-domain mutation (I23A) with that of the well-characterized I59T β-domain variant. We also investigated the effect of a mutation that has minor effects on native-state stability at the domain interface (I56V) and compared it with that of a variant with similar stability within the C-helix (I89V). We show that when variants have similar reduced native-state stabilities, the location of the mutation (I23A versus I59T) is crucial to the native-state dynamics, with the α-domain mutation having a significantly lower ability to populate transient intermediate species under physiologically relevant conditions. Interestingly, the mutation at the interface (I56V) has a greater effect in facilitating the formation of transient intermediate species at elevated temperatures compared with the variants containing α-domain mutations, even though this mutation results in only minor changes to the native-state stability of lysozyme. These findings reveal that the location of specific mutations is an important factor in determining the native-state dynamical properties of human lysozyme in the context of its propensity to populate the aggregation-prone transient intermediate species associated with pathogenic amyloid formation.
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影响因子：
2.9
作者：
Canet, D;Sunde, M;Dobson, CM
通讯作者：
Dobson, CM
影响因子：
13.8
作者：
Dobson, CM
通讯作者：
Dobson, CM
影响因子：
2.9
作者：
Mombelli, E;Afshar, M;Lange, R
通讯作者：
Lange, R
DOI：
10.1093/protein/12.10.841
发表时间：
1999-10-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Funahashi, J;Takano, K;Yutani, K
通讯作者：
Yutani, K
DOI：
10.1021/jp403425z
发表时间：
2013-10-24
期刊：
The journal of physical chemistry. B
影响因子：
--
作者：
De Genst E;Chan PH;Pardon E;Hsu SD;Kumita JR;Christodoulou J;Menzer L;Chirgadze DY;Robinson CV;Muyldermans S;Matagne A;Wyns L;Dobson CM;Dumoulin M
通讯作者：
Dumoulin M