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Mechanistic and structural analysis of a family 31 ?-glycosidase and its lycosyl-enzyme intermediate

Mechanistic and structural analysis of a family 31 ?-glycosidase and its lycosyl-enzyme intermediate
Mechanistic and structural analysis of a family 31 ?-glycosidase and its lycosyl-enzyme intermediate
We have determined the first structure of a family 31 ?-glycosidase, that of YicI from Escherichia coli, both free and trapped as a 5-fluoroxylopyranosyl-enzyme intermediate via reaction with 5-fluoro-?-D-xylopyranosyl fluoride. Our 2.2-Å resolution structure shows an intimately associated hexamer with structural elements from several monomers converging at each of the six active sites. Our kinetic and mass spectrometry analyses verified several of the features observed in our structural data, including a covalent linkage from the carboxylate side chain of the identified nucleophile Asp416 to C-1 of the sugar ring. Structure-based sequence comparison of YicI with the mammalian ?-glucosidases lysosomal ?-glucosidase and sucrase-isomaltase predicts a high level of structural similarity and provides a foundation for understanding the various mutations of these enzymes that elicit human disease.
0021-9258
2105-2115
Lovering, Andrew L.
836e794a-429e-4af7-80d0-20d1631355d4
Lee, Seung Seo
ee34fa26-5fb6-48c8-80c2-1f13ec4ccceb
Kim, Young-Wan
c5baeacc-14b4-452e-969e-d9f2132b5e32
Withers, Stephen G.
2fa507a8-6772-41a0-97a0-a33f443bbcc1
Strynadka, Natalie C.
b4848e23-a016-462a-aacb-70b9d142f3a7
Hughes, Howard
3aeab9d6-9d11-4ecb-b238-3bc4e075d7dd
Lovering, Andrew L.
836e794a-429e-4af7-80d0-20d1631355d4
Lee, Seung Seo
ee34fa26-5fb6-48c8-80c2-1f13ec4ccceb
Kim, Young-Wan
c5baeacc-14b4-452e-969e-d9f2132b5e32
Withers, Stephen G.
2fa507a8-6772-41a0-97a0-a33f443bbcc1
Strynadka, Natalie C.
b4848e23-a016-462a-aacb-70b9d142f3a7
Hughes, Howard
3aeab9d6-9d11-4ecb-b238-3bc4e075d7dd

Lovering, Andrew L., Lee, Seung Seo, Kim, Young-Wan, Withers, Stephen G., Strynadka, Natalie C. and Hughes, Howard (2005) Mechanistic and structural analysis of a family 31 ?-glycosidase and its lycosyl-enzyme intermediate. The Journal of Biological Chemistry, 280 (3), 2105-2115. (doi:10.1074/jbc.M410468200).

Record type: Article

Abstract

We have determined the first structure of a family 31 ?-glycosidase, that of YicI from Escherichia coli, both free and trapped as a 5-fluoroxylopyranosyl-enzyme intermediate via reaction with 5-fluoro-?-D-xylopyranosyl fluoride. Our 2.2-Å resolution structure shows an intimately associated hexamer with structural elements from several monomers converging at each of the six active sites. Our kinetic and mass spectrometry analyses verified several of the features observed in our structural data, including a covalent linkage from the carboxylate side chain of the identified nucleophile Asp416 to C-1 of the sugar ring. Structure-based sequence comparison of YicI with the mammalian ?-glucosidases lysosomal ?-glucosidase and sucrase-isomaltase predicts a high level of structural similarity and provides a foundation for understanding the various mutations of these enzymes that elicit human disease.

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More information

Published date: 21 January 2005
Organisations: Organic Chemistry: SCF

Identifiers

Local EPrints ID: 345655
URI: http://eprints.soton.ac.uk/id/eprint/345655
ISSN: 0021-9258
PURE UUID: e3729f41-520e-4720-abb8-f696ef73abce
ORCID for Seung Seo Lee: ORCID iD orcid.org/0000-0002-8598-3303

Catalogue record

Date deposited: 12 Feb 2013 16:39
Last modified: 26 Nov 2019 01:36

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