Quantification of membrane transporter proteins in human lung and immortalized cell lines using targeted quantitative proteomic analysis by isotope dilution nanoLC-MS/MS
Quantification of membrane transporter proteins in human lung and immortalized cell lines using targeted quantitative proteomic analysis by isotope dilution nanoLC-MS/MS
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DOI：
10.1016/j.jpba.2018.02.044
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发表时间：
2018-05-30
影响因子：
3.4
通讯作者：
Smith, Philip C.
中科院分区：
文献类型：
--
作者：
Fallon, John K.;Houvig, Nicole;Smith, Philip C.
需要关于肺转运蛋白表达的信息来为药物开发和治疗决策提供信息。目前可获得的大部分信息来自文献中报道的半定量基因表达或免疫密度测定研究。本文采用NanoLC-MS/MS （MRM模式）同位素稀释靶向定量蛋白质组学方法，对新鲜人肺膜组分样品和选定的永生化人肺上皮细胞系样品的膜组分中选定的12种转运蛋白进行了定量。在粗膜裂解缓冲液中进行均质，然后对均质液进行差动离心。在肺膜中，我们发现oops是检测到的最高表达的转运蛋白，其次是PEPT2和abc （P-gp和BCRP）。SLC22A转运蛋白（OCTs 2 & 3和OCTN1）也被发现表达。OATP2A1，也被称为前列腺素转运蛋白，是表达最高的转运蛋白，在至少偶尔吸烟的两名受试者中表达较低。一个不吸烟的受试者，其OATP2A1的浓度比下一个最近的浓度高8.4倍，而下一个最近的浓度比任何其他转运蛋白的浓度都高。从基因表达和动物功能研究可知，OATP2A1存在于肺中。这些结果为了解肺中的异种生物配置提供了信息，并显示了与其他组织相比，肺中转运蛋白的独特特征。(C) 2018 Elsevier B.V.版权所有
Information is needed on the expression of transporters in lung to inform drug development and therapeutic decisions. Much of the information currently available is from semiquantitative gene expression or immunometric densitometry studies reported in the literature. NanoLC-MS/MS (MRM mode) isotope dilution targeted quantitative proteomics was used here to quantify twelve selected transporters in fresh human lung membrane fraction samples and in the membrane fraction of selected immortalized human lung epithelial cell line samples. Fractionation was undertaken by homogenization in crude membrane lysis buffer followed by differential centrifugation of the homogenate. In lung membranes we found OATPs to be the most highly expressed transporters of those measured, followed by PEPT2 and ABCs (P-gp & BCRP). SLC22A transporters (OCTs 2 & 3 and OCTN1) were also found to be expressed. OATP2A1, also known as the prostaglandin transporter, was the most highly expressed transporter, being low in two subjects who were at least occasional smokers. One subject, a non-smoker, had an OATP2A1 concentration that was 8.4 times higher than the next nearest concentration, which itself was higher than the concentration of any other transporter. OATP2A1 is known, from gene expression and animal functional studies, to be present in lung. These results inform the understanding of xenobiotic disposition in the lung and show the distinct profile of transporters in lung compared to other tissues. (C) 2018 Elsevier B.V. All rights reserved.