Cloning, characterization and tissue distribution of the rat ATP-binding cassette (ABC) transporter ABC2/ABCA2

L. X. Zhao, Chengji Zhou, A. Tanaka, M. Nakata, T. Hirabayashi, T. Amachi, S. Shioda, K. Ueda, N. Inagaki

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

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 250 kDa 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.

Original languageEnglish (US)
Pages (from-to)865-872
Number of pages8
JournalBiochemical Journal
Volume350
Issue number3
DOIs
StatePublished - Sep 15 2000
Externally publishedYes

Fingerprint

ATP-Binding Cassette Transporters
Cloning
Tissue Distribution
Rats
Organism Cloning
Tissue
Brain
Proteins
Adenosine Triphosphate
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Amino Acids
N,N'-bis(4-azidobenzoyl)cystine
COS Cells
Oligodendroglia
Hemagglutinins
Molecular Cloning
Labeling
In Situ Hybridization
Amino Acid Sequence
Glycoproteins

Keywords

  • Apoptosis
  • Brain
  • Flippase
  • Oligodendrocyte

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cloning, characterization and tissue distribution of the rat ATP-binding cassette (ABC) transporter ABC2/ABCA2. / Zhao, L. X.; Zhou, Chengji; Tanaka, A.; Nakata, M.; Hirabayashi, T.; Amachi, T.; Shioda, S.; Ueda, K.; Inagaki, N.

In: Biochemical Journal, Vol. 350, No. 3, 15.09.2000, p. 865-872.

Research output: Contribution to journalArticle

Zhao, LX, Zhou, C, Tanaka, A, Nakata, M, Hirabayashi, T, Amachi, T, Shioda, S, Ueda, K & Inagaki, N 2000, 'Cloning, characterization and tissue distribution of the rat ATP-binding cassette (ABC) transporter ABC2/ABCA2', Biochemical Journal, vol. 350, no. 3, pp. 865-872. https://doi.org/10.1042/0264-6021:3500865
Zhao, L. X. ; Zhou, Chengji ; Tanaka, A. ; Nakata, M. ; Hirabayashi, T. ; Amachi, T. ; Shioda, S. ; Ueda, K. ; Inagaki, N. / Cloning, characterization and tissue distribution of the rat ATP-binding cassette (ABC) transporter ABC2/ABCA2. In: Biochemical Journal. 2000 ; Vol. 350, No. 3. pp. 865-872.
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