The neutron structure of urate oxidase resolves a long-standing mechanistic conundrum and reveals unexpected changes in protonation.
Oksanen, Esko ; Blakeley, Matthew P ; El-Hajji, Mohamed ; Ryde, Ulf LU
and Budayova-Spano, Monika
(2014)
In PLoS ONE
9(1).
- Abstract
- Urate oxidase transforms uric acid to 5-hydroxyisourate without the help of cofactors, but the catalytic mechanism has remained enigmatic, as the protonation state of the substrate could not be reliably deduced. We have determined the neutron structure of urate oxidase, providing unique information on the proton positions. A neutron crystal structure inhibited by a chloride anion at 2.3 Å resolution shows that the substrate is in fact 8-hydroxyxanthine, the enol tautomer of urate. We have also determined the neutron structure of the complex with the inhibitor 8-azaxanthine at 1.9 Å resolution, showing the protonation states of the K10-T57-H256 catalytic triad. Together with X-ray data and quantum chemical calculations, these structures... (More)
- Urate oxidase transforms uric acid to 5-hydroxyisourate without the help of cofactors, but the catalytic mechanism has remained enigmatic, as the protonation state of the substrate could not be reliably deduced. We have determined the neutron structure of urate oxidase, providing unique information on the proton positions. A neutron crystal structure inhibited by a chloride anion at 2.3 Å resolution shows that the substrate is in fact 8-hydroxyxanthine, the enol tautomer of urate. We have also determined the neutron structure of the complex with the inhibitor 8-azaxanthine at 1.9 Å resolution, showing the protonation states of the K10-T57-H256 catalytic triad. Together with X-ray data and quantum chemical calculations, these structures allow us to identify the site of the initial substrate protonation and elucidate why the enzyme is inhibited by a chloride anion. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4290593
- author
- Oksanen, Esko
; Blakeley, Matthew P
; El-Hajji, Mohamed
; Ryde, Ulf
LU
and Budayova-Spano, Monika
- organization
- publishing date
- 2014
- type
- Contribution to journal
- publication status
- published
- subject
- in
- PLoS ONE
- volume
- 9
- issue
- 1
- article number
- e86651
- publisher
- Public Library of Science (PLoS)
- external identifiers
-
- pmid:24466188
- wos:000330288000129
- scopus:84899805626
- pmid:24466188
- ISSN
- 1932-6203
- DOI
- 10.1371/journal.pone.0086651
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Theoretical Chemistry (S) (011001039)
- id
- 811e7984-445f-403e-b67b-9bde82fc3527 (old id 4290593)
- date added to LUP
- 2016-04-01 13:06:44
- date last changed
- 2023-01-11 08:39:15
@article{811e7984-445f-403e-b67b-9bde82fc3527,
abstract = {{Urate oxidase transforms uric acid to 5-hydroxyisourate without the help of cofactors, but the catalytic mechanism has remained enigmatic, as the protonation state of the substrate could not be reliably deduced. We have determined the neutron structure of urate oxidase, providing unique information on the proton positions. A neutron crystal structure inhibited by a chloride anion at 2.3 Å resolution shows that the substrate is in fact 8-hydroxyxanthine, the enol tautomer of urate. We have also determined the neutron structure of the complex with the inhibitor 8-azaxanthine at 1.9 Å resolution, showing the protonation states of the K10-T57-H256 catalytic triad. Together with X-ray data and quantum chemical calculations, these structures allow us to identify the site of the initial substrate protonation and elucidate why the enzyme is inhibited by a chloride anion.}},
author = {{Oksanen, Esko and Blakeley, Matthew P and El-Hajji, Mohamed and Ryde, Ulf and Budayova-Spano, Monika}},
issn = {{1932-6203}},
language = {{eng}},
number = {{1}},
publisher = {{Public Library of Science (PLoS)}},
series = {{PLoS ONE}},
title = {{The neutron structure of urate oxidase resolves a long-standing mechanistic conundrum and reveals unexpected changes in protonation.}},
url = {{https://lup.lub.lu.se/search/files/3165203/5266644.pdf}},
doi = {{10.1371/journal.pone.0086651}},
volume = {{9}},
year = {{2014}},
}