Analysis of amyloid-like secondary structure in the Cryab-R120G knock-in mouse model of hereditary cataracts by two-dimensional infrared spectroscopy.
Analysis of amyloid-like secondary structure in the Cryab-R120G knock-in mouse model of hereditary cataracts by two-dimensional infrared spectroscopy.
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DOI：
10.1371/journal.pone.0257098
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Andley UP
中科院分区：
文献类型：
--
作者：
Alperstein AM;Molnar KS;Dicke SS;Farrell KM;Makley LN;Zanni MT;Andley UP
α B-晶状体蛋白是一种小的热休克蛋白，在眼透镜中与α A-晶状体蛋白形成异源寡聚复合物。它也广泛分布在整个身体的组织中，并与神经退行性疾病如阿尔茨海默氏症有关，其中它与淀粉样纤维有关。晶状体蛋白可以在白内障中形成无定形聚集体以及结构更复杂的淀粉样原纤维。α B-晶状体蛋白（αB-crystallin，Cryab-R120 G）精氨酸120位突变为甘氨酸（R120 G），在体外可导致高分子量晶状体蛋白聚集并丧失分子伴侣活性，与人类遗传性白内障和肌病有关。表征晶体蛋白聚集体的无定形（非结构化）与高度有序（淀粉样原纤维）性质对于理解它们在疾病中的作用是重要的，并且对于开发白内障的药理学治疗是重要的。我们研究了蛋白质二级结构在野生型（WT）和Cryab-R120 G基因敲入突变小鼠晶状体使用二维红外（2D）光谱，已被用于检测淀粉样蛋白样纤维在人类晶状体和测量紫外线辐射诱导的变化，在猪晶状体。我们的目标是比较该小鼠透镜模型与人晶状体中的聚集蛋白，并评估Cryab-R120 G敲入小鼠模型与一般年龄相关性白内障疾病的蛋白结构相关性。在2 π谱中，与WT小鼠晶状体相比，突变小鼠晶状体中酰胺I对角峰频率红移至较小的波数，这与有序二级结构的增加一致。交叉峰频率和强度表明突变小鼠晶状体中存在淀粉样蛋白。虽然来自2 π光谱的位置和强度的对角和交叉峰变化表明野生型和突变小鼠晶状体之间存在显著的结构差异，但这些差异小于在人晶状体中发现的差异;因此，Cryab-R120 G敲入小鼠晶状体比人晶状体含有更少的淀粉样蛋白样二级结构。2 π光谱研究的结果证实了Cryab-R120 G基因敲入小鼠白内障中淀粉样蛋白二级结构的存在，并支持使用该模型研究年龄相关性白内障。
αB-crystallin is a small heat shock protein that forms a heterooligomeric complex with αA-crystallin in the ocular lens. It is also widely distributed in tissues throughout the body and has been linked with neurodegenerative diseases such as Alzheimer’s, where it is associated with amyloid fibrils. Crystallins can form amorphous aggregates in cataracts as well as more structured amyloid-like fibrils. The arginine 120 to glycine (R120G) mutation in αB-crystallin (Cryab-R120G) results in high molecular weight crystallin protein aggregates and loss of the chaperone activity of the protein in vitro, and it is associated with human hereditary cataracts and myopathy. Characterizing the amorphous (unstructured) versus the highly ordered (amyloid fibril) nature of crystallin aggregates is important in understanding their role in disease and important to developing pharmacological treatments for cataracts. We investigated protein secondary structure in wild-type (WT) and Cryab-R120G knock-in mutant mouse lenses using two-dimensional infrared (2DIR) spectroscopy, which has been used to detect amyloid-like fibrils in human lenses and measure UV radiation-induced changes in porcine lenses. Our goal was to compare the aggregated proteins in this mouse lens model to human lenses and evaluate the protein structural relevance of the Cryab-R120G knock-in mouse model to general age-related cataract disease. In the 2DIR spectra, amide I diagonal peak frequencies were red-shifted to smaller wavenumbers in mutant mouse lenses as compared to WT mouse lenses, consistent with an increase in ordered secondary structure. The cross peak frequency and intensity indicated the presence of amyloid in the mutant mouse lenses. While the diagonal and cross peak changes in location and intensity from the 2DIR spectra indicated significant structural differences between the wild type and mutant mouse lenses, these differences were smaller than those found in human lenses; thus, the Cryab-R120G knock-in mouse lenses contain less amyloid-like secondary structure than human lenses. The results of the 2DIR spectroscopy study confirm the presence of amyloid-like secondary structure in Cryab-R120G knock-in mice with cataracts and support the use of this model to study age-related cataract.
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影响因子：
5.5
作者：
Garvey M;Ecroyd H;Ray NJ;Gerrard JA;Carver JA
通讯作者：
Carver JA
DOI：
10.1073/pnas.96.11.6137
发表时间：
1999-05-25
影响因子：
11.1
作者：
Bova, MP;Yaron, O;Horwitz, J
通讯作者：
Horwitz, J
影响因子：
3.8
作者：
Schmeltz, Margaux;Teulon, Claire;Schanne-Klein, Marie-Claire
通讯作者：
Schanne-Klein, Marie-Claire
DOI：
10.1126/science.aac9145
发表时间：
2015-11-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Makley LN;McMenimen KA;DeVree BT;Goldman JW;McGlasson BN;Rajagopal P;Dunyak BM;McQuade TJ;Thompson AD;Sunahara R;Klevit RE;Andley UP;Gestwicki JE
通讯作者：
Gestwicki JE
影响因子：
3.4
作者：
Lampi, KJ;Ma, ZX;David, LL
通讯作者：
David, LL