Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA 70808, USA.
Free Radic Biol Med. 2010 May 15;48(10):1330-7. doi: 10.1016/j.freeradbiomed.2010.02.012. Epub 2010 Feb 24.
Increased levels of misfolded and damaged proteins occur in response to brain aging and Alzheimer disease (AD), which presumably increase the amount of aggregation-prone proteins via elevations in hydrophobicity. The proteasome is an intracellular protease that degrades oxidized and ubiquitinated proteins, and its function is known to be impaired in response to both aging and AD. In this study we sought to determine the potential for increased levels of protein hydrophobicity occurring in response to aging and AD, to identify the contribution of proteasome inhibition to increased protein hydrophobicity, and last to identify the contribution of ubiquitinated and oxidized proteins to the pool of hydrophobic proteins. In our studies we identified that aging and AD brain exhibited increases in protein hydrophobicity as detected using Bis ANS, with dietary restriction (DR) significantly decreasing age-related increases in protein hydrophobicity. Affinity chromatography purification of hydrophobic proteins from aging and AD brains identified increased levels of oxidized and ubiquitinated proteins in the pool of hydrophobic proteins. Pharmacological inhibition of the proteasome in neurons, but not astrocytes, resulted in an increase in protein hydrophobicity. Taken together, these data indicate that there is a relationship between increased protein oxidation and protein ubiquitination and elevations in protein hydrophobicity within the aging and the AD brain, which may be mediated in part by impaired proteasome activity in neurons. Our studies also suggest a potential role for decreased oxidized and hydrophobic proteins in mediating the beneficial effects of DR.
蛋白质错误折叠和损伤水平的增加发生在脑老化和阿尔茨海默病(AD)的反应中,这可能会通过增加疏水性来增加聚集倾向蛋白的量。蛋白酶体是一种细胞内蛋白酶,可降解氧化和泛素化蛋白,其功能已知会因衰老和 AD 而受损。在这项研究中,我们试图确定蛋白质疏水性增加发生在衰老和 AD 反应中的潜在机制,确定蛋白酶体抑制对蛋白质疏水性增加的贡献,最后确定泛素化和氧化蛋白对疏水性蛋白池的贡献。在我们的研究中,我们发现使用 Bis ANS 检测到,衰老和 AD 大脑中的蛋白质疏水性增加,而饮食限制(DR)显著降低了与年龄相关的蛋白质疏水性增加。从衰老和 AD 大脑中疏水性蛋白的亲和色谱纯化鉴定出疏水性蛋白池中氧化和泛素化蛋白水平增加。蛋白酶体在神经元中的药理学抑制,但不是在星形胶质细胞中,导致蛋白质疏水性增加。总之,这些数据表明,在衰老和 AD 大脑中,蛋白质氧化和蛋白质泛素化与蛋白质疏水性升高之间存在关系,这可能部分是由神经元中蛋白酶体活性受损介导的。我们的研究还表明,减少氧化和疏水性蛋白可能在调节 DR 的有益作用中发挥作用。