Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property

Abstract It remains undeciphered how thermophilic enzymes display enhanced stability at elevated temperatures. Taking l-asparaginase from P. furiosus (PfA) as an example, we combined scattering shapes deduced from small-angle X-ray scattering (SAXS) data at increased temperatures with symmetry mates...

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Autores principales:	Pankaj Sharma, Rachana Tomar, Shivpratap Singh Yadav, Maulik D. Badmalia, Samir Kumar Nath, Ashish, Bishwajit Kundu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/cb4ab17049d744219a90d1b286d8a31c
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spelling	oai:doaj.org-article:cb4ab17049d744219a90d1b286d8a31c2021-12-02T11:43:51ZHeat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property10.1038/s41598-020-78877-z2045-2322https://doaj.org/article/cb4ab17049d744219a90d1b286d8a31c2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78877-zhttps://doaj.org/toc/2045-2322Abstract It remains undeciphered how thermophilic enzymes display enhanced stability at elevated temperatures. Taking l-asparaginase from P. furiosus (PfA) as an example, we combined scattering shapes deduced from small-angle X-ray scattering (SAXS) data at increased temperatures with symmetry mates from crystallographic structures to find that heating caused end-to-end association. The small contact point of self-binding appeared to be enabled by a terminal short β-strand in N-terminal domain, Leu179-Val-Val-Asn182 (LVVN). Interestingly, deletion of this strand led to a defunct enzyme, whereas suplementation of the peptide LVVN to the defunct enzyme restored structural frameworkwith mesophile-type functionality. Crystal structure of the peptide-bound defunct enzyme showed that one peptide ispresent in the same coordinates as in original enzyme, explaining gain-of lost function. A second peptide was seen bound to the protein at a different location suggesting its possible role in substrate-free molecular-association. Overall, we show that the heating induced self-assembly of native shapes of PfA led to an apparent super-stable assembly.Pankaj SharmaRachana TomarShivpratap Singh YadavMaulik D. BadmaliaSamir Kumar NathAshishBishwajit KunduNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Pankaj Sharma Rachana Tomar Shivpratap Singh Yadav Maulik D. Badmalia Samir Kumar Nath Ashish Bishwajit Kundu Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property
description	Abstract It remains undeciphered how thermophilic enzymes display enhanced stability at elevated temperatures. Taking l-asparaginase from P. furiosus (PfA) as an example, we combined scattering shapes deduced from small-angle X-ray scattering (SAXS) data at increased temperatures with symmetry mates from crystallographic structures to find that heating caused end-to-end association. The small contact point of self-binding appeared to be enabled by a terminal short β-strand in N-terminal domain, Leu179-Val-Val-Asn182 (LVVN). Interestingly, deletion of this strand led to a defunct enzyme, whereas suplementation of the peptide LVVN to the defunct enzyme restored structural frameworkwith mesophile-type functionality. Crystal structure of the peptide-bound defunct enzyme showed that one peptide ispresent in the same coordinates as in original enzyme, explaining gain-of lost function. A second peptide was seen bound to the protein at a different location suggesting its possible role in substrate-free molecular-association. Overall, we show that the heating induced self-assembly of native shapes of PfA led to an apparent super-stable assembly.
format	article
author	Pankaj Sharma Rachana Tomar Shivpratap Singh Yadav Maulik D. Badmalia Samir Kumar Nath Ashish Bishwajit Kundu
author_facet	Pankaj Sharma Rachana Tomar Shivpratap Singh Yadav Maulik D. Badmalia Samir Kumar Nath Ashish Bishwajit Kundu
author_sort	Pankaj Sharma
title	Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property
title_short	Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property
title_full	Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property
title_fullStr	Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property
title_full_unstemmed	Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property
title_sort	heat induces end to end repetitive association in p. furiosus l-asparaginase which enables its thermophilic property
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/cb4ab17049d744219a90d1b286d8a31c
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Heat induces end to end repetitive association in P. furiosus l-asparaginase which enables its thermophilic property

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