Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases

Abstract GH19 and GH22 glycoside hydrolases belonging to the lysozyme superfamily have a related structure/function. A highly conserved tryptophan residue, Trp103, located in the binding groove of a GH19 chitinase from moss Bryum coronatum (BcChi-A) appears to have a function similar to that of well...

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Autores principales:	Takuya Nagata, Shoko Shinya, Takayuki Ohnuma, Tamo Fukamizo
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/af857347caee45848bbbbf057bade540
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spelling	oai:doaj.org-article:af857347caee45848bbbbf057bade5402021-12-02T14:16:58ZMulti-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases10.1038/s41598-021-81903-32045-2322https://doaj.org/article/af857347caee45848bbbbf057bade5402021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81903-3https://doaj.org/toc/2045-2322Abstract GH19 and GH22 glycoside hydrolases belonging to the lysozyme superfamily have a related structure/function. A highly conserved tryptophan residue, Trp103, located in the binding groove of a GH19 chitinase from moss Bryum coronatum (BcChi-A) appears to have a function similar to that of well-known Trp62 in GH22 lysozymes. Here, we found that mutation of Trp103 to phenylalanine (W103F) or alanine (W103A) strongly reduced the enzymatic activity of BcChi-A. NMR experiments and the X-ray crystal structure suggested a hydrogen bond between the Trp103 side chain and the -2 sugar. Chitooligosaccharide binding experiments using NMR indicated that the W103F mutation reduced the sugar-binding abilities of nearby amino acid residues (Tyr105/Asn106) in addition to Trp103. This appeared to be derived from enhanced aromatic stacking of Phe103 with Tyr105 induced by disruption of the Trp103 hydrogen bond with the -2 sugar. Since the stacking with Tyr105 was unlikely in W103A, Tyr105/Asn106 of W103A was not so affected as in W103F. However, the W103A mutation appeared to reduce the catalytic potency, resulting in the lowest enzymatic activity in W103A. We concluded that Trp103 does not only interact with the sugar, but also controls other amino acids responsible for substrate binding and catalysis. Trp103 (GH19) and Trp62 (GH22) with such a multi-functionality may be advantageous for enzyme action and conserved in the divergent evolution in the lysozyme superfamily.Takuya NagataShoko ShinyaTakayuki OhnumaTamo FukamizoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takuya Nagata Shoko Shinya Takayuki Ohnuma Tamo Fukamizo Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
description	Abstract GH19 and GH22 glycoside hydrolases belonging to the lysozyme superfamily have a related structure/function. A highly conserved tryptophan residue, Trp103, located in the binding groove of a GH19 chitinase from moss Bryum coronatum (BcChi-A) appears to have a function similar to that of well-known Trp62 in GH22 lysozymes. Here, we found that mutation of Trp103 to phenylalanine (W103F) or alanine (W103A) strongly reduced the enzymatic activity of BcChi-A. NMR experiments and the X-ray crystal structure suggested a hydrogen bond between the Trp103 side chain and the -2 sugar. Chitooligosaccharide binding experiments using NMR indicated that the W103F mutation reduced the sugar-binding abilities of nearby amino acid residues (Tyr105/Asn106) in addition to Trp103. This appeared to be derived from enhanced aromatic stacking of Phe103 with Tyr105 induced by disruption of the Trp103 hydrogen bond with the -2 sugar. Since the stacking with Tyr105 was unlikely in W103A, Tyr105/Asn106 of W103A was not so affected as in W103F. However, the W103A mutation appeared to reduce the catalytic potency, resulting in the lowest enzymatic activity in W103A. We concluded that Trp103 does not only interact with the sugar, but also controls other amino acids responsible for substrate binding and catalysis. Trp103 (GH19) and Trp62 (GH22) with such a multi-functionality may be advantageous for enzyme action and conserved in the divergent evolution in the lysozyme superfamily.
format	article
author	Takuya Nagata Shoko Shinya Takayuki Ohnuma Tamo Fukamizo
author_facet	Takuya Nagata Shoko Shinya Takayuki Ohnuma Tamo Fukamizo
author_sort	Takuya Nagata
title	Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
title_short	Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
title_full	Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
title_fullStr	Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
title_full_unstemmed	Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases
title_sort	multi-functionality of a tryptophan residue conserved in substrate-binding groove of gh19 chitinases
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/af857347caee45848bbbbf057bade540
work_keys_str_mv	AT takuyanagata multifunctionalityofatryptophanresidueconservedinsubstratebindinggrooveofgh19chitinases AT shokoshinya multifunctionalityofatryptophanresidueconservedinsubstratebindinggrooveofgh19chitinases AT takayukiohnuma multifunctionalityofatryptophanresidueconservedinsubstratebindinggrooveofgh19chitinases AT tamofukamizo multifunctionalityofatryptophanresidueconservedinsubstratebindinggrooveofgh19chitinases
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Multi-functionality of a tryptophan residue conserved in substrate-binding groove of GH19 chitinases

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