Expression and functional analyses of novel mutations of ATP-binding cassette transporter-1 in Japanese patients with high-density lipoprotein deficiency

Yoshiharu Nishida, Ken Ichi Hirano, Kosuke Tsukamoto, Makoto Nagano, Chiaki Ikegami, Zhongyan Zhang, Ken ichi Tsujii, Akifumi Matsuyama, Tohru Ohama, Fumihiko Matsuura, Masato Ishigami, Naohiko Sakai, Hisatoyo Hiraoka, Shizuya Yamashita, Yuji Matsuzawa, Makoto Nagano, Mitsuaki Ishihara, Hiroaki Hattori, Toru Egashira, Naoki SakaneYoshihide Yoshida, Kirsten Roomp, Cheryl Wellington, Michael R. Hayden, Susumu Misugi

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26 Citations (Scopus)

Abstract

ATP-binding cassette transporter-1 (ABCA1) gene is mutated in patients with familial high-density lipoprotein deficiency (FHD). In order to know the molecular basis for FHD, we characterized three different ABCA1 mutations associated with FHD (G1158A/A255T, C5946T/R1851X, and A5226G/N1611D) with respect to their expression in the passaged fibroblasts from the patients and in the cells transfected with the mutated cDNAs. Fibroblasts from the all patients showed markedly decreased cholesterol efflux to apolipoprotein (apo)-Al. In the fibroblasts homozygous for G1158A/A255T, the immunoreactive mass of ABCA1 could not be detected, even when stimulated by 9-cis-retinoic acid and 22-R-hydroxycholesterol. In the fibroblasts homozygous for C5946T/R1851X, ABCA1 mRNA was comparable. Because the mutant ABCA1 protein (R1851X) was predicted to lack the epitope for the antibody used, we transfected FLAG-tagged truncated mutant (R1851X/ABCA1-FLAG) cDNA into Cos-7 cells, showing that the mutant protein expression was markedly reduced. The expression of N1611D ABCA1 protein was comparable in both fibroblasts and over-expressing cells, although cholesterol efflux from the cells was markedly reduced. These data indicated that, in the three patients investigated, the abnormalities and dysfunction of ABCA1 occurred at the different levels, providing important information about the expression, regulation, and function of ABCA1.

Original languageEnglish
Pages (from-to)713-721
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume290
Issue number2
DOIs
Publication statusPublished - 2002

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Nishida, Y., Hirano, K. I., Tsukamoto, K., Nagano, M., Ikegami, C., Zhang, Z., Tsujii, K. I., Matsuyama, A., Ohama, T., Matsuura, F., Ishigami, M., Sakai, N., Hiraoka, H., Yamashita, S., Matsuzawa, Y., Nagano, M., Ishihara, M., Hattori, H., Egashira, T., ... Misugi, S. (2002). Expression and functional analyses of novel mutations of ATP-binding cassette transporter-1 in Japanese patients with high-density lipoprotein deficiency. Biochemical and Biophysical Research Communications, 290(2), 713-721. https://doi.org/10.1006/bbrc.2001.6219