Aggrelyte-2 promotes protein solubility and decreases lens stiffness through lysine acetylation and disulfide reduction: Implications for treating presbyopia.
Aggrelyte-2 promotes protein solubility and decreases lens stiffness through lysine acetylation and disulfide reduction: Implications for treating presbyopia.
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作者：
晶状体中老化的蛋白质变得越来越聚集和不溶，导致老花眼。在这项研究中，我们研究了aggrelyte - 2 （N，S -二乙酰- L -半胱氨酸甲酯）逆转老化的人类晶状体蛋白的水不溶性和降低培养的人类和小鼠晶状体硬度的能力。用aggrelyte‐2 （500 μM）孵育65-75岁老年性人晶状体的水不溶性蛋白（WI） 24或48小时。缺少S -乙酰基（aggrelyte - 2C）的对照化合物也进行了测试。我们观察到，在用聚集体- 2处理后，WI的溶解度为19%-30%。Aggrelyte‐2C也增加了蛋白质的溶解度，但其效果比Aggrelyte‐2低约1.4倍。与未处理的样品相比，聚合物- 2和聚合物- 2C处理过的样品中蛋白质硫醇含量高1.9 - 4.9倍。LC-MS /MS结果显示，在aggrelyte‐2和aggrelyte‐2C处理的样品中，N ε‐乙酰赖氨酸（AcK）水平为1.5 ~ 2.1 nmol/mg蛋白质和0.6 ~ 0.9 nmol/mg蛋白质。用1.0 mM aggrelyte‐2处理的小鼠（C57BL/6J）晶状体（孵育24小时）和人晶状体（孵育72小时）的硬度显著降低，可溶性蛋白（人晶状体）和蛋白- AcK水平同时增加，而在aggrelyte‐2C‐处理的晶状体中没有观察到这种变化。溶解蛋白的质谱分析显示，AcK在所有结晶蛋白中都存在，但在α‐结晶蛋白中观察到更多。这些结果表明，aggrelyte‐2通过乙酰化和二硫还原增加了蛋白质的溶解度，降低了晶状体的硬度。Aggrelyte‐2可能对治疗人类老花眼有用。老花眼是许多40岁以上人群的主要视力障碍问题。晶状体硬化（变硬），由于蛋白质不溶解和聚集，显著有助于老花眼。Aggrelyte‐2溶解聚集的蛋白质，并通过赖氨酸乙酰化和二硫还原降低晶状体硬度。
Aging proteins in the lens become increasingly aggregated and insoluble, contributing to presbyopia. In this study, we investigated the ability of aggrelyte‐2 (N,S‐diacetyl‐L‐cysteine methyl ester) to reverse the water insolubility of aged human lens proteins and to decrease stiffness in cultured human and mouse lenses. Water‐insoluble proteins (WI) of aged human lenses (65–75 years) were incubated with aggrelyte‐2 (500 μM) for 24 or 48 h. A control compound that lacked the S‐acetyl group (aggrelyte‐2C) was also tested. We observed 19%–30% solubility of WI upon treatment with aggrelyte‐2. Aggrelyte‐2C also increased protein solubility, but its effect was approximately 1.4‐fold lower than that of aggrelyte‐2. The protein thiol contents were 1.9‐ to 4.9‐fold higher in the aggrelyte‐2‐ and aggrelyte‐2C‐treated samples than in the untreated samples. The LC–MS/MS results showed N ε ‐acetyllysine (AcK) levels of 1.5 to 2.1 nmol/mg protein and 0.6 to 0.9 nmol/mg protein in the aggrelyte‐2‐ and aggrelyte‐2C‐treated samples. Mouse (C57BL/6J) lenses (incubated for 24 h) and human lenses (incubated for 72 h) with 1.0 mM aggrelyte‐2 showed significant decreases in stiffness with simultaneous increases in soluble proteins (human lenses) and protein‐AcK levels, and such changes were not observed in aggrelyte‐2C‐treated lenses. Mass spectrometry of the solubilized protein revealed AcK in all crystallins, but more was observed in α‐crystallins. These results suggest that aggrelyte‐2 increases protein solubility and decreases lens stiffness through acetylation and disulfide reduction. Aggrelyte‐2 might be useful in treating presbyopia in humans. Presbyopia is a major vision‐impeding problem for many people over 40. Lens hardening (stiffening), due to protein insolubilization and aggregation, significantly contributes to presbyopia. Aggrelyte‐2 solubilizes aggregated proteins and reduces lens stiffness through lysine acetylation and disulfide reduction.
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DOI：
10.1016/j.bbadis.2020.165998
发表时间：
2021-01-01
期刊：
Biochimica et biophysica acta. Molecular basis of disease
影响因子：
--
作者：
Karnam S;Skiba NP;Rao PV
通讯作者：
Rao PV
影响因子：
3.4
作者：
Hariharapura R;Santhoshkumar P;Krishna Sharma K
通讯作者：
Krishna Sharma K
影响因子：
4.4
作者：
Hains, Peter G.;Truscott, Roger J. W.
通讯作者：
Truscott, Roger J. W.
影响因子：
4.8
作者：
Cobb, BA;Petrash, JM
通讯作者：
Petrash, JM
DOI：
10.1073/pnas.75.8.3618
发表时间：
1978-01-01
影响因子：
11.1
作者：
GARNER, WH;SPECTOR, A
通讯作者：
SPECTOR, A