An old method facing a new challenge: re-visiting housekeeping proteins as internal reference control for neuroscience research.
An old method facing a new challenge: re-visiting housekeeping proteins as internal reference control for neuroscience research.
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DOI：
10.1016/j.lfs.2013.02.014
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发表时间：
2013-04-19
期刊：
影响因子：
6.1
通讯作者：
Shen, Yong
中科院分区：
文献类型：
--
作者：
Li, Rena;Shen, Yong
特异性靶蛋白表达的研究通常通过蛋白质印迹进行，这是一种在神经科学研究中通过特异性抗体测量蛋白质表达的常用方法。管家蛋白用作蛋白质上样的内部对照以及蛋白质印迹分析中的参考。这种做法是基于这样的信念，即管家基因被认为是无处不在的，在每一个组织中组成型表达，并产生正常细胞功能所必需的最低限度的必需转录本。最常用的管家蛋白是β-肌动蛋白、β-微管蛋白和甘油醛3-磷酸脱氢酶（GAPDH）。然而，最近的研究表明，在各种神经病理学事件，如脊髓损伤和阿尔茨海默病中，一些管家基因在mRNA和蛋白质水平上都存在显着变化。管家基因的变化也由各种组织中的非神经元疾病诱导。因此，这些发现提出了一个潜在的问题，是否使用管家蛋白作为目标蛋白分析的内标是一个适当的做法。这个小型审查将集中在（I）神经元和非神经元疾病的影响，实验条件下，和组织特异性的作用改变管家基因，和（II）替代基因和蛋白质表达分析的内部标准。
Study of specific target protein expression is often performed by western blotting, a commonly used method to measure the protein expression in neuroscience research by specific antibodies. Housekeeping proteins are used as an internal control for protein loading as well as reference in the western blotting analysis. This practice is based on the belief that such housekeeping genes are considered to be ubiquitously and constitutively expressed in every tissue and produce the minimal essential transcripts necessary for normal cellular function. The most commonly used housekeeping proteins are β-actin, β-tubulin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). However, recent studies have shown significant variation in some housekeeping genes both at the mRNA and protein levels in various neuropathological events, such as spinal cord injury and Alzheimer's diseases. Changes of housekeeping genes are also induced by non-neuronal diseases in various tissues. Therefore, these discoveries raise a potential concern regarding whether using a housekeeping protein as an internal standard for target protein analysis is an appropriate practice. This mini review will focus on (I) the effects of neuronal and non-neuronal diseases, experimental condition, and tissues-specific roles on alteration of housekeeping genes, and (II) alternative internal standards for gene and protein expression analysis.
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