Quantitative Targeted Absolute Proteomics for 28 Transporters in Brush-Border and Basolateral Membrane Fractions of Rat Kidney.
Quantitative Targeted Absolute Proteomics for 28 Transporters in Brush-Border and Basolateral Membrane Fractions of Rat Kidney.
复制标题
大鼠肾脏刷状缘和基底外侧膜组分中 28 种转运蛋白的定量靶向绝对蛋白质组学。
DOI：
10.1002/jps.24645
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Uchida Y
中科院分区：
文献类型：
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作者：
岡崎翔吾;境太希;谷内出友美;石川稔;橋本祐一;山口卓男;Uchida Y
本研究的目的是确定各种转运蛋白在肾刷状缘膜（BBM）和基底外侧膜（BLM）组分中的绝对蛋白表达水平，以了解不同转运蛋白在每个细胞膜上的平均转运活性的定量差异。制备大鼠肾脏BBM和BLM组分，用胰蛋白酶消化，并使用我们建立的定量靶向绝对蛋白质组学（QTAP）技术对28种转运蛋白和BLM标记物Na+/K+-ATP酶进行同时绝对定量。在BBM组分中，bcrp、urat 1、mate 1、octl 1、mrp 4、mdr 1a和abca 3的蛋白表达水平分别为40.3、22.2、8.90、4.85、4.69、3.22和0.976 fmol/μg蛋白。在BLM组分中，oat 1、oat 3、oct 1、mrp 6和mrp 1的蛋白表达水平分别为10.6、10.2、4.59、0.724和0.271 fmol/μg蛋白。两种组分中abca 2、abca 4、abca 5、abca 12、abcb 4、mrp 5、abcc 9、abcg 1、abcg 5、lat 1、ntcp、pgt、oatp 2b 1、oatp 1b 2、oatp 3a 1和oct 3的表达水平均低于定量限。在这些膜的定量转运蛋白的表达谱，通过QTAP分析确定，应有助于了解个别转运蛋白的肾脏排泄的外源性物质和内源性化合物的贡献。
The purpose of the present study was to determine the absolute protein expression levels of various transporters in renal brush-border membrane (BBM) and basolateral membrane (BLM) fractions, in order to understand the quantitative differences in average transport activities among different transporters at each cellular membrane. BBM and BLM fractions of rat kidney were prepared and digested with trypsin, and simultaneous absolute quantification of 28 transporters and a BLM marker, Na+/K+-ATPase, was performed using our established quantitative-targeted absolute proteomics (QTAP) technique. In BBM fraction, the protein expression levels of bcrp, urat1, mate1, octl1, mrp4, mdr1a, and abca3 were 40.3, 22.2, 8.90, 4.85, 4.69, 3.22, and 0.976 fmol/μg protein, respectively. In BLM fraction, the protein expression levels of oat1, oat3, oct1, mrp6, and mrp1 were 10.6, 10.2, 4.59, 0.724, and 0.271 fmol/μg protein, respectively. The expression levels of abca2, abca4, abca5, abca12, abcb4, mrp5, abcc9, abcg1, abcg5, lat1, ntcp, pgt, oatp2b1, oatp1b2, oatp3a1, and oct3 were under the limit of quantification in both fractions. The quantitative transporter protein expression profiles at these membranes, as determined by QTAP analysis, should be helpful to understand the contributions of individual transporters to renal excretion of xenobiotics and endogenous compounds.