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Probing biophysical sequence constraints within the transmembrane domains of rhodopsin by deep mutational scanning
- Source :
- Science Advances
- Publication Year :
- 2020
- Publisher :
- American Association for the Advancement of Science, 2020.
-
Abstract
- Sequence constraints associated with topological energetics restrict the evolution and mutational tolerance of membrane proteins.<br />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.
- Subjects :
- Models, Molecular
Protein Folding
Rhodopsin
Lipid Bilayers
Biophysics
Gene Expression
Biochemistry
Protein Structure, Secondary
03 medical and health sciences
Protein Domains
Humans
Research Articles
030304 developmental biology
0303 health sciences
Multidisciplinary
biology
Chemistry
Bilayer
030302 biochemistry & molecular biology
Cell Membrane
Solvation
SciAdv r-articles
Recombinant Proteins
Folding (chemistry)
Transmembrane domain
HEK293 Cells
Membrane protein
Solubility
Mutation
biology.protein
Thermodynamics
Sequence space (evolution)
Hydrophobic and Hydrophilic Interactions
Function (biology)
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 23752548
- Volume :
- 6
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Science Advances
- Accession number :
- edsair.doi.dedup.....7181e33c7c3b9c034c2bc6d56f3bf56e