Proteomics-Based Transporter Identification by the PICK Method: Involvement of TM7SF3 and LHFPL6 in Proton-Coupled Organic Cation Antiport at the Blood-Brain Barrier

被引：11

作者：

Kurosawa, Toshiki ^{[1
]}

Tega, Yuma ^{[1
,2
]}

Uchida, Yasuo ^{[3
]}

Higuchi, Kei ^{[4
]}

Tabata, Hidetsugu ^{[5
]}

Sumiyoshi, Takaaki ^{[6
]}

Kubo, Yoshiyuki ^{[1
]}

Terasaki, Tetsuya ^{[3
,7
]}

Deguchi, Yoshiharu ^{[1
]}

机构：

[1] Teikyo Univ, Fac Pharma Sci, Lab Drug Disposit & Pharmacokinet, Tokyo 1738605, Japan

[2] Univ Kentucky, Coll Pharm, Dept Pharmaceut Sci, Lexington, KY 40536 USA

[3] Tohoku Univ, Grad Sch Pharmaceut Sci, Div Membrane Transport & Drug Targeting, Sendai, Miyagi 9808578, Japan

[4] Tokyo Univ Pharm & Life Sci, Sch Pharm, Dept Biopharmaceut, Tokyo 1920392, Japan

[5] Teikyo Univ, Med Chem Lab, Fac Pharma Sci, Tokyo 1738605, Japan

[6] Kansai Univ, Dept Life Sci & Biotechnol, Fac Chem Mat & Bioengn, Osaka 5648680, Japan

[7] Univ Eastern Finland, Sch Pharm, Fac Hlth Sci, FI-70210 Kuopio, Finland

来源：

PHARMACEUTICS | 2022年 / 14卷 / 08期

基金：

日本学术振兴会;

关键词：

proton-coupled organic cation antiporter; blood-brain barrier; photo-affinity labeling; proteomics; SWATH-MS (sequential window acquisition of all theoretical-mass spectra); NICOTINE TRANSPORT; IN-VITRO; RAT; H-1-ANTAGONIST; OXYCODONE; MEMBER; CELLS; LIVER;

D O I：

10.3390/pharmaceutics14081683

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

A proton-coupled organic cation (H+/OC) antiporter working at the blood-brain barrier (BBB) in humans and rodents is thought to be a promising candidate for the efficient delivery of cationic drugs to the brain. Therefore, it is important to identify the molecular entity that exhibits this activity. Here, for this purpose, we established the Proteomics-based Identification of transporter by Crosslinking substrate in Keyhole (PICK) method, which combines photo-affinity labeling with comprehensive proteomics analysis using SWATH-MS. Using preselected criteria, the PICK method generated sixteen candidate proteins. From these, knockdown screening in hCMEC/D3 cells, an in vitro BBB model, identified two proteins, TM7SF3 and LHFPL6, as candidates for the H+/OC antiporter. We synthesized a novel H+/OC antiporter substrate for functional analysis of TM7SF3 and LHFPL6 in hCMEC/D3 cells and HEK293 cells. The results suggested that both TM7SF3 and LHFPL6 are components of the H+/OC antiporter.

引用

页数：19

共 6 条

[1] Involvement of a proton-coupled organic cation antiporter in the blood-brain barrier transport of amantadine
Suzuki, Toyofumi
Aoyama, Takahiko
Suzuki, Naoto
Kobayashi, Masaru
Fukami, Toshiro
Matsumoto, Yoshiaki
Tomono, Kazuo
BIOPHARMACEUTICS & DRUG DISPOSITION, 2016, 37 (06) : 323 - 335
[2] Involvement of Proton-Coupled Organic Cation Antiporter in Human Blood-Brain Barrier Transport of Mesoridazine and Metoclopramide
Debori, Yasuyuki
Igari, Tomoko
Nakakariya, Masanori
Hirabayashi, Hideki
Aoyama, Kazunobu
Amano, Nobuyuki
Kurosawa, Toshiki
Kubo, Yoshiyuki
Deguchi, Yoshiharu
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 2024, 47 (10) : 1662 - 1667
[3] Involvement of Proton-Coupled Organic Cation Antiporter in Varenicline Transport at Blood-Brain Barrier of Rats and in Human Brain Capillary Endothelial Cells
Kurosawa, Toshiki
Higuchi, Kei
Okura, Takashi
Kobayashi, Kazumasa
Kusuhara, Hiroyuki
Deguchi, Yoshiharu
JOURNAL OF PHARMACEUTICAL SCIENCES, 2017, 106 (09) : 2576 - 2582
[4] Memantine transport by a proton-coupled organic cation antiporter in hCMEC/D3 cells, an in vitro human blood-brain barrier model
Higuchi, Kei
Kitamura, Atsushi
Okura, Takashi
Deguchi, Yoshiharu
DRUG METABOLISM AND PHARMACOKINETICS, 2015, 30 (02) : 182 - 187
[5] Naltrexone Transport by a Proton-Coupled Organic Cation Antiporter in hCMEC/D3 Cells, an in Vitro Human Blood-Brain Barrier Model
Kitamura, Atsushi
Higuchi, Kei
Kurosawa, Toshiki
Okura, Takashi
Kubo, Yoshiyuki
Deguchi, Yoshiharu
BIOLOGICAL & PHARMACEUTICAL BULLETIN, 2022, 45 (10) : 1585 - 1589
[6] Functional expression of a proton-coupled organic cation (H+/OC) antiporter in human brain capillary endothelial cell line hCMEC/D3, a human blood-brain barrier model
Shimomura K.
Okura T.
Kato S.
Couraud P.-O.
Schermann J.-M.
Terasaki T.
Deguchi Y.
Fluids and Barriers of the CNS, 10 (1)

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