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Dimerisation induced formation of the active site and the identification of three metal sites in EAL-phosphodiesterases

Dimerisation induced formation of the active site and the identification of three metal sites in EAL-phosphodiesterases
Dimerisation induced formation of the active site and the identification of three metal sites in EAL-phosphodiesterases

The bacterial second messenger cyclic di-3',5'-guanosine monophosphate (c-di-GMP) is a key regulator of bacterial motility and virulence. As high levels of c-di-GMP are associated with the biofilm lifestyle, c-di-GMP hydrolysing phosphodiesterases (PDEs) have been identified as key targets to aid development of novel strategies to treat chronic infection by exploiting biofilm dispersal. We have studied the EAL signature motif-containing phosphodiesterase domains from the Pseudomonas aeruginosa proteins PA3825 (PA3825EAL) and PA1727 (MucREAL). Different dimerisation interfaces allow us to identify interface independent principles of enzyme regulation. Unlike previously characterised two-metal binding EAL-phosphodiesterases, PA3825EAL in complex with pGpG provides a model for a third metal site. The third metal is positioned to stabilise the negative charge of the 5'-phosphate, and thus three metals could be required for catalysis in analogy to other nucleases. This newly uncovered variation in metal coordination may provide a further level of bacterial PDE regulation.

Journal Article
2045-2322
42166
Bellini, Dom
39c25764-e748-4639-9df2-197e74083002
Horrell, Sam
a9a13833-811f-4932-971c-1bd2e36c3ad2
Hutchin, Andrew
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Phippen, Curtis W
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Strange, Richard W
120909e0-1357-4fdb-9f1d-a0daadf9e414
Cai, Yuming
b6d250da-396a-4807-aa74-0fd3b034ba03
Wagner, Armin
4ffa8ad8-0190-47bf-b5da-8da47c8dada6
Webb, Jeremy S
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Tews, Ivo
9117fc5e-d01c-4f8d-a734-5b14d3eee8dd
Walsh, Martin A
c51d771b-68b4-4c84-8906-650fc25bdad3
Bellini, Dom
39c25764-e748-4639-9df2-197e74083002
Horrell, Sam
a9a13833-811f-4932-971c-1bd2e36c3ad2
Hutchin, Andrew
5659cf31-f52e-451e-9cf0-4e786924d1ae
Phippen, Curtis W
82362439-962f-43e1-89a9-20b2b6bb4b88
Strange, Richard W
120909e0-1357-4fdb-9f1d-a0daadf9e414
Cai, Yuming
b6d250da-396a-4807-aa74-0fd3b034ba03
Wagner, Armin
4ffa8ad8-0190-47bf-b5da-8da47c8dada6
Webb, Jeremy S
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Tews, Ivo
9117fc5e-d01c-4f8d-a734-5b14d3eee8dd
Walsh, Martin A
c51d771b-68b4-4c84-8906-650fc25bdad3

Bellini, Dom, Horrell, Sam, Hutchin, Andrew, Phippen, Curtis W, Strange, Richard W, Cai, Yuming, Wagner, Armin, Webb, Jeremy S, Tews, Ivo and Walsh, Martin A (2017) Dimerisation induced formation of the active site and the identification of three metal sites in EAL-phosphodiesterases. Scientific Reports, 7, 42166. (doi:10.1038/srep42166).

Record type: Article

Abstract

The bacterial second messenger cyclic di-3',5'-guanosine monophosphate (c-di-GMP) is a key regulator of bacterial motility and virulence. As high levels of c-di-GMP are associated with the biofilm lifestyle, c-di-GMP hydrolysing phosphodiesterases (PDEs) have been identified as key targets to aid development of novel strategies to treat chronic infection by exploiting biofilm dispersal. We have studied the EAL signature motif-containing phosphodiesterase domains from the Pseudomonas aeruginosa proteins PA3825 (PA3825EAL) and PA1727 (MucREAL). Different dimerisation interfaces allow us to identify interface independent principles of enzyme regulation. Unlike previously characterised two-metal binding EAL-phosphodiesterases, PA3825EAL in complex with pGpG provides a model for a third metal site. The third metal is positioned to stabilise the negative charge of the 5'-phosphate, and thus three metals could be required for catalysis in analogy to other nucleases. This newly uncovered variation in metal coordination may provide a further level of bacterial PDE regulation.

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Accepted/In Press date: 5 January 2017
e-pub ahead of print date: 10 February 2017
Published date: 10 February 2017
Keywords: Journal Article

Identifiers

Local EPrints ID: 422350
URI: https://eprints.soton.ac.uk/id/eprint/422350
ISSN: 2045-2322
PURE UUID: 04ef346d-0eb5-40e3-ba21-100403bec9d6
ORCID for Jeremy S Webb: ORCID iD orcid.org/0000-0003-2068-8589
ORCID for Ivo Tews: ORCID iD orcid.org/0000-0002-4704-1139

Catalogue record

Date deposited: 20 Jul 2018 16:31
Last modified: 14 Mar 2019 01:41

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Contributors

Author: Dom Bellini
Author: Sam Horrell
Author: Andrew Hutchin
Author: Curtis W Phippen
Author: Richard W Strange
Author: Yuming Cai
Author: Armin Wagner
Author: Jeremy S Webb ORCID iD
Author: Ivo Tews ORCID iD
Author: Martin A Walsh

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