Soluble variants of human recombinant glutaminyl cyclase.

Recombinant human Glutaminyl Cyclase expressed in E. coli is produced as inclusion bodies. Lack of glycosylation is the main origin of its accumulation in insoluble aggregates. Mutation of single isolated hydrophobic amino acids into negative amino acids was not able to circumvent inclusion bodies f...

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Autores principales: Cristiana Castaldo, Silvia Ciambellotti, Raquel de Pablo-Latorre, Daniela Lalli, Valentina Porcari, Paola Turano
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Publicado: Public Library of Science (PLoS) 2013
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Acceso en línea:https://doaj.org/article/08b72f1f483a4db7a361c7e1338f8eac
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spelling oai:doaj.org-article:08b72f1f483a4db7a361c7e1338f8eac2021-11-18T08:59:25ZSoluble variants of human recombinant glutaminyl cyclase.1932-620310.1371/journal.pone.0071657https://doaj.org/article/08b72f1f483a4db7a361c7e1338f8eac2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23977104/?tool=EBIhttps://doaj.org/toc/1932-6203Recombinant human Glutaminyl Cyclase expressed in E. coli is produced as inclusion bodies. Lack of glycosylation is the main origin of its accumulation in insoluble aggregates. Mutation of single isolated hydrophobic amino acids into negative amino acids was not able to circumvent inclusion bodies formation. On the contrary, substitution with carboxyl-terminal residues of two or three aromatic residues belonging to extended hydrophobic patches on the protein surface provided soluble but still active forms of the protein. These mutants could be expressed in isotopically enriched forms for NMR studies and the maximal attainable concentration was sufficient for the acquisition of (1)H-(15)N HSQC spectra that represent the starting point for future drug development projects targeting Alzheimer's disease.Cristiana CastaldoSilvia CiambellottiRaquel de Pablo-LatorreDaniela LalliValentina PorcariPaola TuranoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 8, p e71657 (2013)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Cristiana Castaldo
Silvia Ciambellotti
Raquel de Pablo-Latorre
Daniela Lalli
Valentina Porcari
Paola Turano
Soluble variants of human recombinant glutaminyl cyclase.
description Recombinant human Glutaminyl Cyclase expressed in E. coli is produced as inclusion bodies. Lack of glycosylation is the main origin of its accumulation in insoluble aggregates. Mutation of single isolated hydrophobic amino acids into negative amino acids was not able to circumvent inclusion bodies formation. On the contrary, substitution with carboxyl-terminal residues of two or three aromatic residues belonging to extended hydrophobic patches on the protein surface provided soluble but still active forms of the protein. These mutants could be expressed in isotopically enriched forms for NMR studies and the maximal attainable concentration was sufficient for the acquisition of (1)H-(15)N HSQC spectra that represent the starting point for future drug development projects targeting Alzheimer's disease.
format article
author Cristiana Castaldo
Silvia Ciambellotti
Raquel de Pablo-Latorre
Daniela Lalli
Valentina Porcari
Paola Turano
author_facet Cristiana Castaldo
Silvia Ciambellotti
Raquel de Pablo-Latorre
Daniela Lalli
Valentina Porcari
Paola Turano
author_sort Cristiana Castaldo
title Soluble variants of human recombinant glutaminyl cyclase.
title_short Soluble variants of human recombinant glutaminyl cyclase.
title_full Soluble variants of human recombinant glutaminyl cyclase.
title_fullStr Soluble variants of human recombinant glutaminyl cyclase.
title_full_unstemmed Soluble variants of human recombinant glutaminyl cyclase.
title_sort soluble variants of human recombinant glutaminyl cyclase.
publisher Public Library of Science (PLoS)
publishDate 2013
url https://doaj.org/article/08b72f1f483a4db7a361c7e1338f8eac
work_keys_str_mv AT cristianacastaldo solublevariantsofhumanrecombinantglutaminylcyclase
AT silviaciambellotti solublevariantsofhumanrecombinantglutaminylcyclase
AT raqueldepablolatorre solublevariantsofhumanrecombinantglutaminylcyclase
AT danielalalli solublevariantsofhumanrecombinantglutaminylcyclase
AT valentinaporcari solublevariantsofhumanrecombinantglutaminylcyclase
AT paolaturano solublevariantsofhumanrecombinantglutaminylcyclase
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