Structural basis for substrate specificity of mammalian neuraminidases.

The removal of sialic acid (Sia) residues from glycoconjugates in vertebrates is mediated by a family of neuraminidases (sialidases) consisting of Neu1, Neu2, Neu3 and Neu4 enzymes. The enzymes play distinct physiological roles, but their ability to discriminate between the types of linkages connect...

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Autores principales:	Victoria Smutova, Amgad Albohy, Xuefang Pan, Elena Korchagina, Taeko Miyagi, Nicolai Bovin, Christopher W Cairo, Alexey V Pshezhetsky
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d530e95bd42943088afd4f242855bc27
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spelling	oai:doaj.org-article:d530e95bd42943088afd4f242855bc272021-11-25T06:00:37ZStructural basis for substrate specificity of mammalian neuraminidases.1932-620310.1371/journal.pone.0106320https://doaj.org/article/d530e95bd42943088afd4f242855bc272014-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0106320https://doaj.org/toc/1932-6203The removal of sialic acid (Sia) residues from glycoconjugates in vertebrates is mediated by a family of neuraminidases (sialidases) consisting of Neu1, Neu2, Neu3 and Neu4 enzymes. The enzymes play distinct physiological roles, but their ability to discriminate between the types of linkages connecting Sia and adjacent residues and between the identity and arrangement of the underlying sugars has never been systematically studied. Here we analyzed the specificity of neuraminidases by studying the kinetics of hydrolysis of BODIPY-labeled substrates containing common mammalian sialylated oligosaccharides: 3'Sia-LacNAc, 3'SiaLac, SiaLex, SiaLea, SiaLec, 6'SiaLac, and 6'SiaLacNAc. We found significant differences in substrate specificity of the enzymes towards the substrates containing α2,6-linked Sia, which were readily cleaved by Neu3 and Neu1 but not by Neu4 and Neu2. The presence of a branching 2-Fuc inhibited Neu2 and Neu4, but had almost no effect on Neu1 or Neu3. The nature of the sugar residue at the reducing end, either glucose (Glc) or N-acetyl-D-glucosamine (GlcNAc) had only a minor effect on all neuraminidases, whereas core structure (1,3 or 1,4 bond between D-galactose (Gal) and GlcNAc) was found to be important for Neu4 strongly preferring β3 (core 1) to β4 (core 2) isomer. Neu3 and Neu4 were in general more active than Neu1 and Neu2, likely due to their preference for hydrophobic substrates. Neu2 and Neu3 were examined by molecular dynamics to identify favorable substrate orientations in the binding sites and interpret the differences in their specificities. Finally, using knockout mouse models, we confirmed that the substrate specificities observed in vitro were recapitulated in enzymes found in mouse brain tissues. Our data for the first time provide evidence for the characteristic substrate preferences of neuraminidases and their ability to discriminate between distinct sialoside targets.Victoria SmutovaAmgad AlbohyXuefang PanElena KorchaginaTaeko MiyagiNicolai BovinChristopher W CairoAlexey V PshezhetskyPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 9, p e106320 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Victoria Smutova Amgad Albohy Xuefang Pan Elena Korchagina Taeko Miyagi Nicolai Bovin Christopher W Cairo Alexey V Pshezhetsky Structural basis for substrate specificity of mammalian neuraminidases.
description	The removal of sialic acid (Sia) residues from glycoconjugates in vertebrates is mediated by a family of neuraminidases (sialidases) consisting of Neu1, Neu2, Neu3 and Neu4 enzymes. The enzymes play distinct physiological roles, but their ability to discriminate between the types of linkages connecting Sia and adjacent residues and between the identity and arrangement of the underlying sugars has never been systematically studied. Here we analyzed the specificity of neuraminidases by studying the kinetics of hydrolysis of BODIPY-labeled substrates containing common mammalian sialylated oligosaccharides: 3'Sia-LacNAc, 3'SiaLac, SiaLex, SiaLea, SiaLec, 6'SiaLac, and 6'SiaLacNAc. We found significant differences in substrate specificity of the enzymes towards the substrates containing α2,6-linked Sia, which were readily cleaved by Neu3 and Neu1 but not by Neu4 and Neu2. The presence of a branching 2-Fuc inhibited Neu2 and Neu4, but had almost no effect on Neu1 or Neu3. The nature of the sugar residue at the reducing end, either glucose (Glc) or N-acetyl-D-glucosamine (GlcNAc) had only a minor effect on all neuraminidases, whereas core structure (1,3 or 1,4 bond between D-galactose (Gal) and GlcNAc) was found to be important for Neu4 strongly preferring β3 (core 1) to β4 (core 2) isomer. Neu3 and Neu4 were in general more active than Neu1 and Neu2, likely due to their preference for hydrophobic substrates. Neu2 and Neu3 were examined by molecular dynamics to identify favorable substrate orientations in the binding sites and interpret the differences in their specificities. Finally, using knockout mouse models, we confirmed that the substrate specificities observed in vitro were recapitulated in enzymes found in mouse brain tissues. Our data for the first time provide evidence for the characteristic substrate preferences of neuraminidases and their ability to discriminate between distinct sialoside targets.
format	article
author	Victoria Smutova Amgad Albohy Xuefang Pan Elena Korchagina Taeko Miyagi Nicolai Bovin Christopher W Cairo Alexey V Pshezhetsky
author_facet	Victoria Smutova Amgad Albohy Xuefang Pan Elena Korchagina Taeko Miyagi Nicolai Bovin Christopher W Cairo Alexey V Pshezhetsky
author_sort	Victoria Smutova
title	Structural basis for substrate specificity of mammalian neuraminidases.
title_short	Structural basis for substrate specificity of mammalian neuraminidases.
title_full	Structural basis for substrate specificity of mammalian neuraminidases.
title_fullStr	Structural basis for substrate specificity of mammalian neuraminidases.
title_full_unstemmed	Structural basis for substrate specificity of mammalian neuraminidases.
title_sort	structural basis for substrate specificity of mammalian neuraminidases.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/d530e95bd42943088afd4f242855bc27
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_version_	1718414309197873152

Structural basis for substrate specificity of mammalian neuraminidases.

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