Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.

We present a combined in vitro/in silico study to determine the molecular origin of the selectivity of [Formula: see text]-tocopherol transfer protein ([Formula: see text]-TTP) towards [Formula: see text]-tocopherol. Molecular dynamics simulations combined to free energy perturbation calculations pr...

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Autores principales:	Rachel E Helbling, Walter Aeschimann, Fabio Simona, Achim Stocker, Michele Cascella
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e04ddf7bcf24488fb18c3a6d1a593ada
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spelling	oai:doaj.org-article:e04ddf7bcf24488fb18c3a6d1a593ada2021-11-18T08:09:00ZEngineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.1932-620310.1371/journal.pone.0049195https://doaj.org/article/e04ddf7bcf24488fb18c3a6d1a593ada2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23152872/?tool=EBIhttps://doaj.org/toc/1932-6203We present a combined in vitro/in silico study to determine the molecular origin of the selectivity of [Formula: see text]-tocopherol transfer protein ([Formula: see text]-TTP) towards [Formula: see text]-tocopherol. Molecular dynamics simulations combined to free energy perturbation calculations predict a binding free energy for [Formula: see text]-tocopherol to [Formula: see text]-TTP 8.26[Formula: see text]2.13 kcal mol[Formula: see text] lower than that of [Formula: see text]-tocopherol. Our calculations show that [Formula: see text]-tocopherol binds to [Formula: see text]-TTP in a significantly distorted geometry as compared to that of the natural ligand. Variations in the hydration of the binding pocket and in the protein structure are found as well. We propose a mutation, A156L, which significantly modifies the selectivity properties of [Formula: see text]-TTP towards the two tocopherols. In particular, our simulations predict that A156L binds preferentially to [Formula: see text]-tocopherol, with striking structural similarities to the wild-type-[Formula: see text]-tocopherol complex. The affinity properties are confirmed by differential scanning fluorimetry as well as in vitro competitive binding assays. Our data indicate that residue A156 is at a critical position for determination of the selectivity of [Formula: see text]-TTP. The engineering of TTP mutants with modulating binding properties can have potential impact at industrial level for easier purification of single tocopherols from vitamin E mixtures coming from natural oils or synthetic processes. Moreover, the identification of a [Formula: see text]-tocopherol selective TTP offers the possibility to challenge the hypotheses for the evolutionary development of a mechanism for [Formula: see text]-tocopherol selection in omnivorous animals.Rachel E HelblingWalter AeschimannFabio SimonaAchim StockerMichele CascellaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 11, p e49195 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rachel E Helbling Walter Aeschimann Fabio Simona Achim Stocker Michele Cascella Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.
description	We present a combined in vitro/in silico study to determine the molecular origin of the selectivity of [Formula: see text]-tocopherol transfer protein ([Formula: see text]-TTP) towards [Formula: see text]-tocopherol. Molecular dynamics simulations combined to free energy perturbation calculations predict a binding free energy for [Formula: see text]-tocopherol to [Formula: see text]-TTP 8.26[Formula: see text]2.13 kcal mol[Formula: see text] lower than that of [Formula: see text]-tocopherol. Our calculations show that [Formula: see text]-tocopherol binds to [Formula: see text]-TTP in a significantly distorted geometry as compared to that of the natural ligand. Variations in the hydration of the binding pocket and in the protein structure are found as well. We propose a mutation, A156L, which significantly modifies the selectivity properties of [Formula: see text]-TTP towards the two tocopherols. In particular, our simulations predict that A156L binds preferentially to [Formula: see text]-tocopherol, with striking structural similarities to the wild-type-[Formula: see text]-tocopherol complex. The affinity properties are confirmed by differential scanning fluorimetry as well as in vitro competitive binding assays. Our data indicate that residue A156 is at a critical position for determination of the selectivity of [Formula: see text]-TTP. The engineering of TTP mutants with modulating binding properties can have potential impact at industrial level for easier purification of single tocopherols from vitamin E mixtures coming from natural oils or synthetic processes. Moreover, the identification of a [Formula: see text]-tocopherol selective TTP offers the possibility to challenge the hypotheses for the evolutionary development of a mechanism for [Formula: see text]-tocopherol selection in omnivorous animals.
format	article
author	Rachel E Helbling Walter Aeschimann Fabio Simona Achim Stocker Michele Cascella
author_facet	Rachel E Helbling Walter Aeschimann Fabio Simona Achim Stocker Michele Cascella
author_sort	Rachel E Helbling
title	Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.
title_short	Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.
title_full	Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.
title_fullStr	Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.
title_full_unstemmed	Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.
title_sort	engineering tocopherol selectivity in α-ttp: a combined in vitro/in silico study.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/e04ddf7bcf24488fb18c3a6d1a593ada
work_keys_str_mv	AT rachelehelbling engineeringtocopherolselectivityinattpacombinedinvitroinsilicostudy AT walteraeschimann engineeringtocopherolselectivityinattpacombinedinvitroinsilicostudy AT fabiosimona engineeringtocopherolselectivityinattpacombinedinvitroinsilicostudy AT achimstocker engineeringtocopherolselectivityinattpacombinedinvitroinsilicostudy AT michelecascella engineeringtocopherolselectivityinattpacombinedinvitroinsilicostudy
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Engineering tocopherol selectivity in α-TTP: a combined in vitro/in silico study.

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