Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2484307 | Journal of Pharmaceutical Sciences | 2016 | 6 Pages |
Abstract
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.
Keywords
SRM/MRMBBMABCSLCBLMHPLC (high-performance/pressure liquid chromatography)LC-MS/MSsolute transporterssolute carrierMembrane transportDrug transportABC transportersMembrane transporterRenal excretionBBBMass spectrometryLiquid chromatography-tandem mass spectrometryBrush-border membraneBasolateral membraneBlood-brain barrierProteomicsProteinsATP-binding cassette
Related Topics
Health Sciences
Pharmacology, Toxicology and Pharmaceutical Science
Drug Discovery
Authors
Yasuo Uchida, Takafumi Toyohara, Sumio Ohtsuki, Yoshinori Moriyama, Takaaki Abe, Tetsuya Terasaki,