Kinetic control in protein folding for light chain amyloidosis and the differential effects of somatic mutations.
Kinetic control in protein folding for light chain amyloidosis and the differential effects of somatic mutations.
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DOI：
10.1016/j.jmb.2013.10.016
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发表时间：
2014-01-23
影响因子：
5.6
通讯作者：
Ramirez-Alvarado M
中科院分区：
文献类型：
--
作者：
Blancas-Mejía LM;Tischer A;Thompson JR;Tai J;Wang L;Auton M;Ramirez-Alvarado M
轻链淀粉样变性是一种破坏性疾病，其中免疫球蛋白轻链形成淀粉样纤维，导致器官功能障碍和死亡。先前的研究表明，蛋白质的热力学稳定性与轻链淀粉样变性中某些蛋白质的淀粉样蛋白形成倾向之间存在直接相关性。在这里，我们调查的影响，体细胞突变的蛋白质稳定性和体外原纤维形成的单和双恢复突变体的蛋白质AL-103相比，野生型生殖系对照蛋白。的扫描速率依赖性和滞后的热展开和重折叠中观察到的所有蛋白质。这表明，解折叠/重折叠反应的动力学确定与不同的动力学常数的解折叠和重折叠，即使该过程仍然实验可逆。我们对AL-103和AL-103 delP95aIns的结构分析表明，解折叠/重折叠过程与顺式-反式脯氨酰异构化存在动力学耦合。我们的数据显示，脯氨酸95a（AL-103 delP95aIns）的缺失（其去除了患者蛋白AL-103中存在的反式顺式二脯氨酸基序）导致热力学稳定性的显著增加和相对于AL-103的原纤维形成动力学的显著延迟。原纤维的形成是pH依赖性的;所有蛋白质在pH 2下形成原纤维;当pH增加到pH 7时，反应变得更慢和更随机。基于这些结果，我们提出，除了热力学稳定性，动力学稳定性（可能受顺式脯氨酸95a的存在影响），在AL-103淀粉样蛋白原纤维形成过程中起着重要作用。
Light chain amyloidosis is a devastating disease where immunoglobulin light chains form amyloid fibrils, resulting in organ dysfunction and death. Previous studies have shown a direct correlation between the protein thermodynamic stability and the propensity for amyloid formation for some proteins involved in light chain amyloidosis. Here we investigate the effect of somatic mutations on protein stability and in vitro fibril formation of single and double restorative mutants of the protein AL-103 compared to the wild type germline control protein. A scan rate dependence and hysteresis in the thermal unfolding and refolding was observed for all proteins. This indicates that the unfolding/refolding reaction is kinetically determined with different kinetic constants for unfolding and refolding even though the process remains experimentally reversible. Our structural analysis of AL-103 and AL-103 delP95aIns suggests a kinetic coupling of the unfolding/refolding process with cis-trans prolyl isomerization. Our data reveal that the deletion of Proline 95a (AL-103 delP95aIns), which removes the trans cis diproline motif present in the patient protein AL-103, results in a dramatic increment in the thermodynamic stability and a significant delay in fibril formation kinetics with respect to AL-103. Fibril formation is pH dependent; all proteins form fibrils at pH 2; reactions become slower and more stochastic as the pH increases up to pH 7. Based on these results, we propose that in addition to thermodynamic stability, kinetic stability (possibly influenced by the presence of cis Proline 95a), plays a major role in the AL-103 amyloid fibril formation process.
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DOI：
10.1038/nsb750
发表时间：
2002-02-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Fasshauer, D;Antonin, W;Jahn, R
通讯作者：
Jahn, R
DOI：
10.1073/pnas.96.7.3590
发表时间：
1999-03-30
影响因子：
11.1
作者：
Chiti, F;Webster, P;Dobson, CM
通讯作者：
Dobson, CM
影响因子：
2.9
作者：
BOLEN, DW;SANTORO, MM
通讯作者：
SANTORO, MM
影响因子：
4.8
作者：
DiCostanzo, Ara Celi;Thompson, James R.;Ramirez-Alvarado, Marina
通讯作者：
Ramirez-Alvarado, Marina
影响因子：
4.8
作者：
Kim, YS;Wall, JS;Carpenter, JF
通讯作者：
Carpenter, JF