Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Nov;181(22):4593-4609.
doi: 10.1111/bph.16508. Epub 2024 Aug 2.

In silico and biological analyses of missense variants of the human biliary efflux transporter ABCC2: effects of novel rare missense variants

Charlotte Kölz  1   2   3 , Fabienne Z Gaugaz  4 , Niklas Handin  4 , Elke Schaeffeler  1   2   3 , Roman Tremmel  1   2 , Stefan Winter  1   2 , Kathrin Klein  1   2 , Ulrich M Zanger  1   2 , Per Artursson  4 , Matthias Schwab  1   2   3   5 , Anne T Nies  1   2   3
Affiliations
Free article

In silico and biological analyses of missense variants of the human biliary efflux transporter ABCC2: effects of novel rare missense variants

Charlotte Kölz et al. Br J Pharmacol. 2024 Nov.
Free article

Abstract

Background and purpose: The ATP-dependent biliary efflux transporter ABCC2, also known as multidrug resistance protein 2 (MRP2), is essential for the cellular disposition and detoxification of various xenobiotics including drugs as well as endogenous metabolites. Common functionally relevant ABCC2 genetic variants significantly alter drug responses and contribute to side effects. The aim of this study was to determine functional consequences of rare variants identified in subjects with European ancestry using in silico tools and in vitro analyses.

Experimental approach: Targeted next-generation sequencing of the ABCC2 gene was used to identify novel variants in European subjects (n = 143). Twenty-six in silico tools were used to predict functional consequences. For biological validation, transport assays were carried out with membrane vesicles prepared from cell lines overexpressing the newly identified ABCC2 variants and estradiol β-glucuronide and carboxydichlorofluorescein as the substrates.

Key results: Three novel rare ABCC2 missense variants were identified (W227R, K402T, V489F). Twenty-five in silico tools predicted W227R as damaging and one as potentially damaging. Prediction of functional consequences was not possible for K402T and V489F and for the common linked variants V1188E/C1515Y. Characterisation in vitro showed increased function of W227R, V489F and V1188E/C1515Y for both substrates, whereas K402T function was only increased for carboxydichlorofluorescein.

Conclusion and implications: In silico tools were unable to accurately predict the substrate-dependent increase in function of ABCC2 missense variants. In vitro biological studies are required to accurately determine functional activity to avoid misleading consequences for drug therapy.

Keywords: ABCC2; MRP2; functional assay; in silico prediction; membrane vesicles; rare missense variant; transport studies.

PubMed Disclaimer

References

REFERENCES

    1. Abramson, J., Adler, J., Dunger, J., Evans, R., Green, T., Pritzel, A., Ronneberger, O., Willmore, L., Ballard, A. J., Bambrick, J., Bodenstein, S. W., Evans, D. A., Hung, C.‐C., O'Neill, M., Reiman, D., Tunyasuvunakool, K., Wu, Z., Žemgulytė, A., Arvaniti, E., … Jumper, J. M. (2024). Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature, 630, 493–500. https://doi.org/10.1038/s41586-024-07487-w
    1. Alexander, S. P. H., Fabbro, D., Kelly, E., Mathie, A. A., Peters, J. A., Veale, E. L., Armstrong, J. F., Faccenda, E., Harding, S. D., Davies, J. A., Amarosi, L., Anderson, C. M. H., Beart, P. M., Broer, S., Dawson, P. A., Gyimesi, G., Hagenbuch, B., Hammond, J. R., Hancox, J. C., … Verri, T. (2023). The Concise Guide to PHARMACOLOGY 2023/24: Transporters. British Journal of Pharmacology, 180(Suppl 2), S374–S469. https://doi.org/10.1111/bph.16182
    1. Alexander, S. P. H., Kelly, E., Mathie, A. A., Peters, J. A., Veale, E. L., Armstrong, J. F., Buneman, O. P., Faccenda, E., Harding, S. D., Spedding, M., Cidlowski, J. A., Fabbro, D., Davenport, A. P., Striessnig, J., Davies, J. A., Ahlers‐Dannen, K. E., Alqinyah, M., Arumugam, T. V., Bodle, C., … Zolghadri, Y. (2023). The Concise Guide to PHARMACOLOGY 2023/24: Introduction and Other Protein Targets. British Journal of Pharmacology, 180, S1–S22. https://doi.org/10.1111/bph.16176
    1. Alexander, S. P. H., Roberts, R. E., Broughton, B. R. S., Sobey, C. G., George, C. H., Stanford, S. C., Cirino, G., Docherty, J. R., Giembycz, M. A., Hoyer, D., Insel, P. A., Izzo, A. A., Ji, Y., MacEwan, D. J., Mangum, J., Wonnacott, S., & Ahluwalia, A. (2018). Goals and practicalities of immunoblotting and immunohistochemistry: A guide for submission to the British Journal of Pharmacology. British Journal of Pharmacology, 175, 407–411. https://doi.org/10.1111/bph.14112
    1. Arlanov, R., Lang, T., Jedlitschky, G., Schaeffeler, E., Ishikawa, T., Schwab, M., & Nies, A. T. (2016). Functional characterization of common protein variants in the efflux transporter ABCC11 and identification of T546M as functionally‐damaging variant. The Pharmacogenomics Journal, 16(2), 193–201. https://doi.org/10.1038/tpj.2015.27

Substances

LinkOut - more resources