Quantitative Targeted Absolute Proteomics for 28 Transporters in Brush-Border and Basolateral Membrane Fractions of Rat Kidney

Yasuo Uchida, Takafumi Toyohara, Sumio Ohtsuki, Yoshinori Moriyama, Takaaki Abe, Tetsuya Terasaki

Research output: Contribution to journalArticle

11 Citations (Scopus)

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.

Original languageEnglish
JournalJournal of Pharmaceutical Sciences
DOIs
Publication statusAccepted/In press - 2015

Fingerprint

Microvilli
Proteomics
Kidney
Membranes
Proteins
Xenobiotics
Trypsin

Keywords

  • ABC transporters
  • Drug transport
  • HPLC (high-performance/pressure liquid chromatography)
  • Mass spectrometry
  • Membrane transport
  • Membrane transporter
  • Proteins
  • Proteomics
  • Renal excretion
  • Solute transporters

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

Quantitative Targeted Absolute Proteomics for 28 Transporters in Brush-Border and Basolateral Membrane Fractions of Rat Kidney. / Uchida, Yasuo; Toyohara, Takafumi; Ohtsuki, Sumio; Moriyama, Yoshinori; Abe, Takaaki; Terasaki, Tetsuya.

In: Journal of Pharmaceutical Sciences, 2015.

Research output: Contribution to journalArticle

Uchida, Yasuo ; Toyohara, Takafumi ; Ohtsuki, Sumio ; Moriyama, Yoshinori ; Abe, Takaaki ; Terasaki, Tetsuya. / Quantitative Targeted Absolute Proteomics for 28 Transporters in Brush-Border and Basolateral Membrane Fractions of Rat Kidney. In: Journal of Pharmaceutical Sciences. 2015.
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AB - 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.

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KW - Proteins

KW - Proteomics

KW - Renal excretion

KW - Solute transporters

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