2. Academic Validation
  3. Characterization of human ATP-binding cassette protein subfamily D reconstituted into proteoliposomes

Characterization of human ATP-binding cassette protein subfamily D reconstituted into proteoliposomes

  • Biochem Biophys Res Commun. 2018 Feb 19;496(4):1122-1127. doi: 10.1016/j.bbrc.2018.01.153.
Takumi Okamoto 1,  Kosuke Kawaguchi 2,  Shiro Watanabe 3,  Rina Agustina 1,  Toshiki Ikejima 1,  Keisuke Ikeda 4,  Minoru Nakano 4,  Masashi Morita 1,  Tsuneo Imanaka 5
Affiliations

Affiliations

  • 1 Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
  • 2 Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan. Electronic address: kkawa@pha.u-toyama.ac.jp.
  • 3 Institute of Natural Medicine, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
  • 4 Laboratory of Biointerface Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan.
  • 5 Department of Biological Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama, 930-0194, Japan; Faculty of Pharmaceutical Sciences, Hiroshima International University, Hirokoshinkai 5-1-1, Kure, 737-0112, Japan.
PMID: 29397936 DOI: 10.1016/j.bbrc.2018.01.153
Abstract

In mammals, four ATP-binding cassette (ABC) proteins belonging to subfamily D have been identified. ABCD1‒3 are located on peroxisomal membrane and play an important role in the transportation of various fatty acid-CoA derivatives, including very long chain fatty acid-CoA, into peroxisomes. ABCD4 is located on lysosomal membrane and is suggested to be involved in the transport of vitamin B12 from lysosomes to the cytosol. However, the precise transport mechanism by which these ABC transporters facilitate the import or export of substrate has yet to be well elucidated. In this study, the overexpression of human ABCD1‒4 in the methylotrophic yeast Pichia pastoris and a purification procedure were developed. The detergent-solubilized proteins were reconstituted into liposomes. ABCD1‒4 displayed stable ATPase activity, which was inhibited by AlF3. Furthermore, ABCD1‒4 were found to possess an equal levels of acyl-CoA thioesterase activity. Proteoliposomes is expected to be an aid in the further biochemical characterization of ABCD transporters.

Keywords

ABC protein subfamily D; ATPase; Acyl-CoA thioesterase; Proteoliposome; Purification.

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