title: Mutational scanning reveals the determinants of protein insertion and association energetics in the plasma membrane.
abstract: Insertion of helix-forming segments into the membrane and their association determines the structure, function, and expression levels of all plasma membrane proteins. However, systematic and reliable quantification of membrane-protein energetics has been challenging. We developed a deep mutational scanning method to monitor the effects of hundreds of point mutations on helix insertion and self-association within the bacterial inner membrane. The assay quantifies insertion energetics for all natural amino acids at 27 positions across the membrane, revealing that the hydrophobicity of biological membranes is significantly higher than appreciated. We further quantitate the contributions to membrane-protein insertion from positively charged residues at the cytoplasm-membrane interface and reveal large and unanticipated differences among these residues. Finally, we derive comprehensive mutational landscapes in the membrane domains of Glycophorin A and the ErbB2 oncogene, and find that insertion and self-association are strongly coupled in receptor homodimers.
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Assaf
authors.initials: A
authors.lastname: Elazar
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Weinstein
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Ido
authors.initials: I
authors.lastname: Biran
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Yearit
authors.initials: Y
authors.lastname: Fridman
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Eitan
authors.initials: E
authors.lastname: Bibi
authors.affiliation: Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Sarel Jacob
authors.identifier: http://orcid.org/0000-0002-6831-3770
authors.initials: SJ
authors.lastname: Fleishman
mesh_terms: D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D004252:DNA Mutational Analysis; D006021:Glycophorins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D011485:Protein Binding; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D006021:Glycophorins; D008565:Membrane Proteins; D050505:Mutant Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D001618:beta-Lactamases; C508053:ERBB2 protein, human
keywords: <i>e. coli</i>; biophysics; deep sequencing; erbB2; high throughput; membrane; rosetta; structural biology
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