Probing biophysical sequence constraints within the transmembrane domains of rhodopsin by deep mutational scanning.

Abstract:

:Membrane proteins must balance the sequence constraints associated with folding and function against the hydrophobicity required for solvation within the bilayer. We recently found the expression and maturation of rhodopsin are limited by the hydrophobicity of its seventh transmembrane domain (TM7), which contains polar residues that are essential for function. On the basis of these observations, we hypothesized that rhodopsin's expression should be less tolerant of mutations in TM7 relative to those within hydrophobic TM domains. To test this hypothesis, we used deep mutational scanning to compare the effects of 808 missense mutations on the plasma membrane expression of rhodopsin in HEK293T cells. Our results confirm that a higher proportion of mutations within TM7 (37%) decrease rhodopsin's plasma membrane expression relative to those within a hydrophobic TM domain (TM2, 25%). These results in conjunction with an evolutionary analysis suggest solvation energetics likely restricts the evolutionary sequence space of polar TM domains.

journal_name

Sci Adv

journal_title

Science advances

authors

Penn WD,McKee AG,Kuntz CP,Woods H,Nash V,Gruenhagen TC,Roushar FJ,Chandak M,Hemmerich C,Rusch DB,Meiler J,Schlebach JP

doi

10.1126/sciadv.aay7505

subject

Has Abstract

pub_date

2020-03-04 00:00:00

pages

eaay7505

issue

10

issn

2375-2548

pii

aay7505

journal_volume

6

pub_type

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