Quantitative Targeted Absolute Proteomics of Transporters and Pharmacoproteomics-Based Reconstruction of P-Glycoprotein Function in Mouse Small Intestine.
The purpose of this study was to investigate whether a pharmacokinetic model integrating in vitro mdr1a efflux activity (which we previously reported) with in vitro/in vivo differences in protein expression level can reconstruct intestinal mdr1a function. In situ intestinal permeability-surface area product ratio between wild-type and mdr1a/1b (-/-) mice is one of the parameters used to describe intestinal mdr1a function. The reconstructed ratios of six mdr1a substrates (dexamethasone, digoxin, loperamide, quinidine, verapamil, vinblastine) and one nonsubstrate (diazepam) were consistent with the observed values reported by Adachi et al. within 2.1-fold difference. Thus, intestinal mdr1a function can be reconstructed by our pharmacoproteomic modeling approach. Furthermore, we evaluated regional differences in protein expression levels of mouse intestinal transporters. Sixteen (mdr1a, mrp4, bcrp, abcg5, abcg8, glut1, 4f2hc, sglt1, lat2, pept1, mct1, slc22a18, ostβ, villin1, Na(+)/K(+)-ATPase, γ-gtp) out of 46 target molecules were detected by employing our established quantitative targeted absolute proteomics technique. The protein expression amounts of mdr1a and bcrp increased progressively from duodenum to ileum. Sglt1, lat2, and 4f2hc were highly expressed in jejunum and ileum. Mct1 and ostβ were highly expressed in ileum. The quantitative expression profiles established here should be helpful to understand and predict intestinal transporter functions.
Division of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University , 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Takanori
T
Akazawa
Division of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University , 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Yasuo
Y
Uchida
Division of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University , 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Masanori
M
Tachikawa
Department of Pharmaceutical Microbiology, Faculty of Life Sciences, Kumamoto University , 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan.
Sumio
S
Ohtsuki
Division of Membrane Transport and Drug Targeting, Graduate School of Pharmaceutical Sciences, Tohoku University , 6-3 Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Tetsuya
T
Terasaki
D018435:ATP Binding Cassette Transporter, Subfamily B / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D003907:Dexamethasone / Q000493:pharmacokinetics; D004077:Digoxin / Q000493:pharmacokinetics; D004386:Duodenum / Q000378:metabolism; D007082:Ileum / Q000378:metabolism; D007408:Intestinal Absorption; D007421:Intestine, Small / Q000378:metabolism*; D007583:Jejunum / Q000378:metabolism; D008139:Loperamide / Q000493:pharmacokinetics; D051379:Mice; D018345:Mice, Knockout; D040901:Proteomics / Q000379:methods*; D011802:Quinidine / Q000493:pharmacokinetics; D014700:Verapamil / Q000493:pharmacokinetics; D014747:Vinblastine / Q000493:pharmacokinetics; D000097202:ATP-Binding Cassette Sub-Family B Member 4
D018435:ATP Binding Cassette Transporter, Subfamily B; D003907:Dexamethasone; D004077:Digoxin; D008139:Loperamide; D011802:Quinidine; D014700:Verapamil; D014747:Vinblastine; D000097202:ATP-Binding Cassette Sub-Family B Member 4; C095122:multidrug resistance protein 3
P-glycoprotein/multidrug resistance protein 1a; drug absorption; pharmacoproteomics; protein quantification; quantitative targeted absolute proteomics; small intestine; transporter