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2-hydroxy-6-keto-nona-2,4-diene 1,9-dioic acid 5,6-hydrolase: evidence from O-18 isotope exchange for gem-diol intermediate

2-hydroxy-6-keto-nona-2,4-diene 1,9-dioic acid 5,6-hydrolase: evidence from O-18 isotope exchange for gem-diol intermediate
2-hydroxy-6-keto-nona-2,4-diene 1,9-dioic acid 5,6-hydrolase: evidence from O-18 isotope exchange for gem-diol intermediate
The mechanism-based inactivation and subsequent identification of the nucleophilic residue using mass spectrometry have been successfully applied and used to identify the active-site nucleophile in numerous ?-glycosidases, as illustrated using C. fimi exoglycanase. Evidence for a covalent glycosyl-enzyme intermediate has come from X-ray crystallographic analysis of trapped complexes, the first being that of the trapped fluoroglycosyl-enzyme intermediate of Cex. The crystal structure of the trapped fluorocellobiosyl-enzyme complex for Cex has provided useful insights into catalysis and the roles of specific residues at the active site. In addition, information about the conformation of the natural sugar in the covalently bound state and the interactions at the active site was obtained using a mutant form of Cex.
alpha/beta-hydrolase-fold, 2-hydroxymuconic semialdehyde hydrolase, c-c hydrolase, 2, 3-dihydroxyphenylpropionate 1, 2-dioxygenase, catecholdioxygenases, pseudomonas-putida, purification, mechanism, degradation, enzymes
0076-6879
106-118
Bugg, Timothy D.H.
300f9ac5-0185-438a-a930-9bf22c08ddd5
Fleming, Sarah M.
c2f6e815-7b3a-4a70-855b-2490baf5f929
Robertson, Thomas A.
c9bed402-93dc-445b-961e-3b048d67dd2c
Langley, G. John
7ac80d61-b91d-4261-ad17-255f94ea21ea
Bugg, Timothy D.H.
300f9ac5-0185-438a-a930-9bf22c08ddd5
Fleming, Sarah M.
c2f6e815-7b3a-4a70-855b-2490baf5f929
Robertson, Thomas A.
c9bed402-93dc-445b-961e-3b048d67dd2c
Langley, G. John
7ac80d61-b91d-4261-ad17-255f94ea21ea

Bugg, Timothy D.H., Fleming, Sarah M., Robertson, Thomas A. and Langley, G. John (2002) 2-hydroxy-6-keto-nona-2,4-diene 1,9-dioic acid 5,6-hydrolase: evidence from O-18 isotope exchange for gem-diol intermediate. Methods in Enzymology, 354, 106-118. (doi:10.1016/S0076-6879(02)54008-8).

Record type: Article

Abstract

The mechanism-based inactivation and subsequent identification of the nucleophilic residue using mass spectrometry have been successfully applied and used to identify the active-site nucleophile in numerous ?-glycosidases, as illustrated using C. fimi exoglycanase. Evidence for a covalent glycosyl-enzyme intermediate has come from X-ray crystallographic analysis of trapped complexes, the first being that of the trapped fluoroglycosyl-enzyme intermediate of Cex. The crystal structure of the trapped fluorocellobiosyl-enzyme complex for Cex has provided useful insights into catalysis and the roles of specific residues at the active site. In addition, information about the conformation of the natural sugar in the covalently bound state and the interactions at the active site was obtained using a mutant form of Cex.

Full text not available from this repository.

More information

Published date: 2002
Keywords: alpha/beta-hydrolase-fold, 2-hydroxymuconic semialdehyde hydrolase, c-c hydrolase, 2, 3-dihydroxyphenylpropionate 1, 2-dioxygenase, catecholdioxygenases, pseudomonas-putida, purification, mechanism, degradation, enzymes

Identifiers

Local EPrints ID: 19689
URI: http://eprints.soton.ac.uk/id/eprint/19689
ISSN: 0076-6879
PURE UUID: 9a6fbc4a-cb08-4295-bc03-e3c23ecf82c2
ORCID for G. John Langley: ORCID iD orcid.org/0000-0002-8323-7235

Catalogue record

Date deposited: 16 Feb 2006
Last modified: 07 Oct 2020 01:34

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