Bridging non-overlapping reads illuminates high-order epistasis between distal protein sites in a GPCR.

Abstract:

:Epistasis emerges when the effects of an amino acid depend on the identities of interacting residues. This phenomenon shapes fitness landscapes, which have the power to reveal evolutionary paths and inform evolution of desired functions. However, there is a need for easily implemented, high-throughput methods to capture epistasis particularly at distal sites. Here, we combine deep mutational scanning (DMS) with a straightforward data processing step to bridge reads in distal sites within genes (BRIDGE). We use BRIDGE, which matches non-overlapping reads to their cognate templates, to uncover prevalent epistasis within the binding pocket of a human G protein-coupled receptor (GPCR) yielding variants with 4-fold greater affinity to a target ligand. The greatest functional improvements in our screen result from distal substitutions and substitutions that are deleterious alone. Our results corroborate findings of mutational tolerance in GPCRs, even in conserved motifs, but reveal inherent constraints restricting tolerated substitutions due to epistasis.

journal_name

Nat Commun

journal_title

Nature communications

authors

Yoo JI,Daugherty PS,O'Malley MA

doi

10.1038/s41467-020-14495-7

subject

Has Abstract

pub_date

2020-02-04 00:00:00

pages

690

issue

1

issn

2041-1723

pii

10.1038/s41467-020-14495-7

journal_volume

11

pub_type

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