2022
DOI: 10.7554/elife.76903
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Determinants of trafficking, conduction, and disease within a K+ channel revealed through multiparametric deep mutational scanning

Abstract: A longstanding goal in protein science and clinical genetics is to develop quantitative models of sequence, structure, and function relationships and delineate the mechanisms by which 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 7,429 single residue missense mutations into the Inward Rectifier K+ channel Kir2.1 and determine how this affects f… Show more

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Cited by 34 publications
(34 citation statements)
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References 102 publications
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“…The FCYENE is in the distal C terminus in non-folding critical regions meaning mutations here likely solely impact ER-export. In contrast, the SY motif interacts directly with the hydrophobic core so SY variants will additionally suffer dramatic folding deficits (Donghui Ma et al 2011;Coyote-Maestas et al 2022;Li et al 2016). With DIMPLE, we can confirm existing phenotypes within known trafficking motifs and in less understood proteins could discover new trafficking motifs and their boundaries.…”
Section: Dimple Libraries Allow Access To Unexplored Sequence Space R...mentioning
confidence: 92%
See 1 more Smart Citation
“…The FCYENE is in the distal C terminus in non-folding critical regions meaning mutations here likely solely impact ER-export. In contrast, the SY motif interacts directly with the hydrophobic core so SY variants will additionally suffer dramatic folding deficits (Donghui Ma et al 2011;Coyote-Maestas et al 2022;Li et al 2016). With DIMPLE, we can confirm existing phenotypes within known trafficking motifs and in less understood proteins could discover new trafficking motifs and their boundaries.…”
Section: Dimple Libraries Allow Access To Unexplored Sequence Space R...mentioning
confidence: 92%
“…Cell lines were generated as in (Coyote-Maestas et al 2022;Matreyek et al 2020): prior to transfection, libraries were cloned into a landing pad vector containing a BxB1-compatible attB recombination site using BsmBI mediated golden gate cloning. We kept track of transformation efficiency to maintain library diversity that was at least 100x the size of a given library.…”
Section: Cell Line Generation and Cell Culturementioning
confidence: 99%
“…3D and Fig. 4, Additional file 3: Fig S10) [34]. Deletions are commonly used by biochemists in an ad hoc fashion to identify important motifs within proteins.…”
Section: Kir21 Surface Expressionmentioning
confidence: 99%
“…Limitations exist in our imaging of OCTN2-tagged variants, where we were unable to quantify colocalization between GFP (OCTN2) and the cell membrane stain due to software limitations and thus instead provided a qualitative classification of cellular membrane localization. As an alternative to high-content imaging, FACS with an antibody against an extracellular domain of OCTN2 or an inserted epitope tag ( 49 ) could be employed to determine membrane localization with more precision.…”
Section: Discussionmentioning
confidence: 99%