An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.

An intimate interaction between a pair of amino acids, a tyrosine and glycine on neighboring β-strands, has been previously reported to be important for the structural stability of autotransporters. Here, we show that the conservation of this interacting pair extends to nearly all major families of...

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Main Authors: Marcin Michalik, Marcella Orwick-Rydmark, Michael Habeck, Vikram Alva, Thomas Arnold, Dirk Linke
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC5542473?pdf=render
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spelling doaj-0ceb679875ca45818a1d4df7b61e7f442020-11-24T21:38:22ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01128e018201610.1371/journal.pone.0182016An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.Marcin MichalikMarcella Orwick-RydmarkMichael HabeckVikram AlvaThomas ArnoldDirk LinkeAn intimate interaction between a pair of amino acids, a tyrosine and glycine on neighboring β-strands, has been previously reported to be important for the structural stability of autotransporters. Here, we show that the conservation of this interacting pair extends to nearly all major families of outer membrane β-barrel proteins, which are thought to have originated through duplication events involving an ancestral ββ hairpin. We analyzed the function of this motif using the prototypical outer membrane protein OmpX. Stopped-flow fluorescence shows that two folding processes occur in the millisecond time regime, the rates of which are reduced in the tyrosine mutant. Folding assays further demonstrate a reduction in the yield of folded protein for the mutant compared to the wild-type, as well as a reduction in thermal stability. Taken together, our data support the idea of an evolutionarily conserved 'folding core' that affects the folding, membrane insertion, and thermal stability of outer membrane protein β-barrels.http://europepmc.org/articles/PMC5542473?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Marcin Michalik
Marcella Orwick-Rydmark
Michael Habeck
Vikram Alva
Thomas Arnold
Dirk Linke
spellingShingle Marcin Michalik
Marcella Orwick-Rydmark
Michael Habeck
Vikram Alva
Thomas Arnold
Dirk Linke
An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
PLoS ONE
author_facet Marcin Michalik
Marcella Orwick-Rydmark
Michael Habeck
Vikram Alva
Thomas Arnold
Dirk Linke
author_sort Marcin Michalik
title An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
title_short An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
title_full An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
title_fullStr An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
title_full_unstemmed An evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
title_sort evolutionarily conserved glycine-tyrosine motif forms a folding core in outer membrane proteins.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2017-01-01
description An intimate interaction between a pair of amino acids, a tyrosine and glycine on neighboring β-strands, has been previously reported to be important for the structural stability of autotransporters. Here, we show that the conservation of this interacting pair extends to nearly all major families of outer membrane β-barrel proteins, which are thought to have originated through duplication events involving an ancestral ββ hairpin. We analyzed the function of this motif using the prototypical outer membrane protein OmpX. Stopped-flow fluorescence shows that two folding processes occur in the millisecond time regime, the rates of which are reduced in the tyrosine mutant. Folding assays further demonstrate a reduction in the yield of folded protein for the mutant compared to the wild-type, as well as a reduction in thermal stability. Taken together, our data support the idea of an evolutionarily conserved 'folding core' that affects the folding, membrane insertion, and thermal stability of outer membrane protein β-barrels.
url http://europepmc.org/articles/PMC5542473?pdf=render
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