A rapid solubility assay of protein domain misfolding for pathogenicity assessment of rare DNA sequence variants.

ABSTRACT:

Purpose

DNA sequencing technology has unmasked a vast number of uncharacterized single-nucleotide variants in disease-associated genes, and efficient methods are needed to determine pathogenicity and enable clinical care.

Methods

We report an E. coli-based solubility assay for assessing the effects of variants on protein domain stability for three disease-associated proteins.

Results

First, we examined variants in the Kv11.1 channel PAS domain (PASD) associated with inherited long QT syndrome type 2 and found that protein solubility correlated well with reported in vitro protein stabilities. A comprehensive solubility analysis of 56 Kv11.1 PASD variants revealed that disruption of membrane trafficking, the dominant loss-of-function disease mechanism, is largely determ

SUBMITTER: Anderson CL 

PROVIDER: S-EPMC7529867 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature