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The G604S-hERG mutation alters the biophysical properties and exerts a dominant-negative effect on expression of hERG channels in HEK293 cells

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Abstract

We have recently identified a missense mutation, G604S, in the human ether-a-go-go related gene (hERG) that results in a malignant phenotype in a full pedigree of a Chinese congenital long QT syndrome (LQTS) family. The present study characterized the pathophysiological consequences of the mutation at the cellular level. Mutant G604S-hERG channels were expressed in HEK293 cells using a lipofectamine method. hERG currents were recorded using the voltage clamp technique. The expression of hERG protein was detected by Western blotting, and the subcellular location of hERG channels in cell was analyzed by confocal microscopy. We found that the G604S mutation did not lead to any expression of detectable currents, which was consistent with Western blotting analysis that the G604S-hERG mutation only expressed a band at 135 kDa. When coexpressed with wild-type (WT)-hERG, G604S-hERG exhibited strong dominant-negative current suppression resulting in decreased current density and altered gating properties of the WT-hERG channel, as well as interference with the trafficking of WT-hERG channel protein. In addition, confocal microscopy demonstrated that G604S-hERG subunits could be inserted into the cell membrane when forming heteromultimeric channels with WT-hERG channel subunits. Our results suggest that G604S mutation causes a loss of function in hERG through a strong dominant-negative effect on WT-hERG channel function that caused by impaired trafficking of WT-hERG channels, and further accentuates this suppression by forming heteromultimeric functional channels with WT-hERG subunits.

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Acknowledgments

This study was supported by grants from National Natural Science Foundation of China (NSFC, numbers 30371571 and 30672209). YMZ thanks the Wellcome Trust for support. CLH and AAG thank the Wellcome Trust, Medical Research Council, and British Heart Foundation for support.

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Authors and Affiliations

  1. Department of Cardiovascular Medicine, First Hospital of Xi’an Jiaotong University, Key Laboratory of Environment and Genes Related to Diseases, Xi’an Jiaotong University, Ministry of Education, No.277 Yanta West Road, Xi’an, Shaanxi, 710061, China

    Jianhua Huo, Yanmin Zhang, Ping Liu, Wenhui Jiang & Aiqun Ma

  2. Medical College of Xi’an Jiaotong University, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, No.277 Yanta West Road, Xi’an, Shaanxi, 710061, China

    Na Huang & Chen Huang

  3. Shaanxi Provincial People’s Hospital, No. 256 West Youyi Road, Xi’an, Shaanxi, 710068, China

    Xueyan Guo

  4. Department of Cardiovascular, Children’s Hospital, No. 69 Xiju Road, Xi’an, Shaanxi, 710002, China

    Nan Zhou

  5. Department of Biochemistry, University of Cambridge, Downing Street, Cambridge, CB2 3EG, UK

    Yanmin Zhang, Andrew Grace & Christopher L. H. Huang

  6. Department of Physiological Laboratory, University of Cambridge, Downing Street, Cambridge, CB2 3EG, UK

    Yanmin Zhang, Andrew Grace & Christopher L. H. Huang

Authors
  1. Jianhua Huo
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  2. Yanmin Zhang
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  3. Na Huang
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  8. Nan Zhou
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  10. Christopher L. H. Huang
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  11. Aiqun Ma
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Correspondence to Aiqun Ma.

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Huo, J., Zhang, Y., Huang, N. et al. The G604S-hERG mutation alters the biophysical properties and exerts a dominant-negative effect on expression of hERG channels in HEK293 cells. Pflugers Arch - Eur J Physiol 456, 917–928 (2008). https://doi.org/10.1007/s00424-008-0454-0

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  • DOI: https://doi.org/10.1007/s00424-008-0454-0

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