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Methods to Examine the Impact of Nonsynonymous SNPs on Protein Degradation and Function of Human ABC Transporter

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Pharmacogenomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1015))

Abstract

Clinical studies have strongly suggested that genetic polymorphisms and/or mutations of certain ATP-binding cassette (ABC) transporter genes might be regarded as significant factors affecting patients’ responses to medication and/or the risk of diseases. In the case of ABCG2, certain single nucleotide polymorphisms (SNPs) in the encoding gene alter the substrate specificity and/or enhance endoplasmic reticulum-associated degradation (ERAD) of the de novo synthesized ABCG2 protein via the ubiquitin-mediated proteasomal proteolysis pathway. Hitherto accumulated clinical data imply that several nonsynonymous SNPs affect the ABCG2-mediated clearance of drugs or cellular metabolites, although some controversies still exist. Therefore, we recently developed high-speed functional screening and ERAD of ABC transporters so as to evaluate the effect of genetic polymorphisms on their function and protein expression levels in vitro. In this chapter we present in vitro experimental methods to elucidate the impact of nonsynonymous SNPs on protein degradation of ABCG2 as well as on its transport function.

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Acknowledgments

The study performed in the authors’ laboratory was supported by the NEDO International Joint Research Grant program “International standardization of functional analysis technology for genetic polymorphisms of drug transporters” as well as a Grant-in-Aid for Scientific Research (A) (No. 18201041) and Grants for Exploratory Research (No. 19659136 and No. 23650619) from the Japanese Society for the Promotion of Science (JSPS).

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Authors and Affiliations

  1. Center for Life Science Technologies, RIKEN, Yokohama, Japan

    Toshihisa Ishikawa

  2. Medical Genetics Division, Shizuoka Cancer Center Research Institute, Shizuoka, Japan

    Kanako Wakabayashi-Nakao

  3. College of Bioscience and Biotechnology, Chubu University, Aichi, Japan

    Hiroshi Nakagawa

Authors
  1. Toshihisa Ishikawa
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  2. Kanako Wakabayashi-Nakao
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  3. Hiroshi Nakagawa
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Editor information

Editors and Affiliations

  1. Institute for Pharmacogenomics, and Individualized Therapy, University of North Carolina, Mason Farm Rd. 120, Chapel Hill, 27599-7361, North Carolina, USA

    Federico Innocenti

  2. , Dept. of Clinical Chemistry (AKC), Erasmus MC, Gravendijkwal 230, Rotterdam, 3015, Netherlands

    Ron H.N. van Schaik

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Ishikawa, T., Wakabayashi-Nakao, K., Nakagawa, H. (2013). Methods to Examine the Impact of Nonsynonymous SNPs on Protein Degradation and Function of Human ABC Transporter. In: Innocenti, F., van Schaik, R. (eds) Pharmacogenomics. Methods in Molecular Biology, vol 1015. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-435-7_15

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  • DOI: https://doi.org/10.1007/978-1-62703-435-7_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-434-0

  • Online ISBN: 978-1-62703-435-7

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