The ABC1 (ABCA) subfamily of the ATP-binding cassette (ABC) transporter superfamily has a structural feature that distinguishes it from other ABC transporters. Here we report the cloning, molecular characterization and tissue distribution of ABC2/ABCA2, which belongs to the ABC1 subfamily. Rat ABC2 is a protein of 2434 amino acids that has 44.5%, 40.0% and 40.8% identity with mouse ABC1/ABCA1, human ABC3/ABCA3 and human ABCR/ABCA4 respectively. Immunoblot analysis showed that proteins of 260 and 250kDa were detected in COS-1 cells transfected with ABC2 having a haemagglutinin tag, while no band was detected in mock-transfected cells. After incubation with N-glycosidase F, the mobilities of the two proteins increased and a single band was detected, suggesting that ABC2 is a glycoprotein. Photoaffinity labelling with 8-azido-[α-32P]ATP confirmed that ATP binds to the ABC2 protein in the presence of Mg2+. RNA blot analysis showed that ABC2 mRNA is most abundant in rat brain. Examination of brain by in situ hybridization determined that ABC2 is expressed at high levels in the white matter, indicating that it is expressed in the oligodendrocytes. ABC2, therefore, is a glycosylated ABC transporter protein, and may play an especially important role in the brain. In addition, the N-terminal 60-amino-acid sequence of the human ABC1, which was missing from previous reports, has been determined.
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Research Article|
September 08 2000
Cloning, characterization and tissue distribution of the rat ATP-binding cassette (ABC) transporter ABC2/ABCA2
Li-Xia ZHAO;
Li-Xia ZHAO
*Department of Physiology, Akita University School of Medicine, 1-1-1, Hondo, Akita 010-8543, Japan
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Cheng-Ji ZHOU;
Cheng-Ji ZHOU
†Department of Anatomy, Showa University School of Medicine, Tokyo 142-8555, Japan
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Arowu TANAKA;
Arowu TANAKA
‡Laboratory of Biochemistry, Division of Applied Life Sciences, Kyoto University Graduate School of Agriculture, Kyoto 606-8502, Japan
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Masanori NAKATA;
Masanori NAKATA
*Department of Physiology, Akita University School of Medicine, 1-1-1, Hondo, Akita 010-8543, Japan
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Takahiro HIRABAYASHI;
Takahiro HIRABAYASHI
§Center for Biotechnology, Showa University, Tokyo 142-8555, Japan
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Teruo AMACHI;
Teruo AMACHI
‡Laboratory of Biochemistry, Division of Applied Life Sciences, Kyoto University Graduate School of Agriculture, Kyoto 606-8502, Japan
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Seiji SHIODA;
Seiji SHIODA
†Department of Anatomy, Showa University School of Medicine, Tokyo 142-8555, Japan
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Kazumitsu UEDA;
Kazumitsu UEDA
‡Laboratory of Biochemistry, Division of Applied Life Sciences, Kyoto University Graduate School of Agriculture, Kyoto 606-8502, Japan
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Nobuya INAGAKI
Nobuya INAGAKI
1
*Department of Physiology, Akita University School of Medicine, 1-1-1, Hondo, Akita 010-8543, Japan
1To whom correspondence should be addressed (e-mail inagaki@med.akita-u.ac.jp).
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Publisher: Portland Press Ltd
Received:
March 14 2000
Revision Received:
June 06 2000
Accepted:
July 12 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2000
2000
Biochem J (2000) 350 (3): 865–872.
Article history
Received:
March 14 2000
Revision Received:
June 06 2000
Accepted:
July 12 2000
Citation
Li-Xia ZHAO, Cheng-Ji ZHOU, Arowu TANAKA, Masanori NAKATA, Takahiro HIRABAYASHI, Teruo AMACHI, Seiji SHIODA, Kazumitsu UEDA, Nobuya INAGAKI; Cloning, characterization and tissue distribution of the rat ATP-binding cassette (ABC) transporter ABC2/ABCA2. Biochem J 15 September 2000; 350 (3): 865–872. doi: https://doi.org/10.1042/bj3500865
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