Determinants of trafficking, conduction, and disease within a K+ channel revealed through multiparametric deep mutational scanning

Willow Coyote-Maestas, David M Nedrud, Yungui He, Daniel Schmidt

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

A long-standing goal in protein science and clinical genetics is to develop quantitative models of sequence, structure, and function relationships to understand how mutations cause disease. Deep mutational scanning (DMS) is a promising strategy to map how amino acids contribute to protein structure and function and to advance clinical variant interpretation. Here, we introduce 7429 single-residue missense mutations into the inward rectifier K + channel Kir2.1 and determine how this affects folding, assembly, and trafficking, as well as regulation by allosteric ligands and ion conduction. Our data provide high-resolution information on a cotranslationally folded biogenic unit, trafficking and quality control signals, and segregated roles of different structural elements in fold stability and function. We show that Kir2.1 surface trafficking mutants are underrepresented in variant effect databases, which has implications for clinical practice. By comparing fitness scores with expert-reviewed variant effects, we can predict the pathogenicity of 'variants of unknown significance' and disease mechanisms of known pathogenic mutations. Our study in Kir2.1 provides a blueprint for how multiparametric DMS can help us understand the mechanistic basis of genetic disorders and the structure-function relationships of proteins.

Original languageEnglish (US)
Article numbere76903
JournaleLife
Volume11
DOIs
StatePublished - May 2022

Bibliographical note

Funding Information:
We are grateful for helpful discussions with Anna Gloyn, James Fraser, Gabbriella Estevam, Eric Greene, the DMS crew, and the rest of the Fraser lab. We also want to acknowledge of the hard from work from the members of the UMN Flow Cytometry Core that enabled us to do these experiments during the COVID-19 pandemic. Rashi Arora especially assisted us in sorting our cells by FACS. We also thank you for taking the time to read our manuscript. This work was supported by the National Institutes of Health (1R01GM136851 to D.S.) and a University of Minnesota Genome Center Illumina S2 grant. W.C.-M. is supported by a National Science Foundation Graduate Research Fellowship and a Howard Hughes Medical Institute Gilliam Fellowship for Advanced Study.

Publisher Copyright:
© 2022, eLife Sciences Publications Ltd. All rights reserved.

Keywords

  • Mutation
  • Mutation, Missense
  • Potassium Channels, Inwardly Rectifying/genetics
  • Proteins/metabolism

PubMed: MeSH publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Journal Article
  • Research Support, N.I.H., Extramural

Fingerprint

Dive into the research topics of 'Determinants of trafficking, conduction, and disease within a K+ channel revealed through multiparametric deep mutational scanning'. Together they form a unique fingerprint.

Cite this