The Rel stringent factor from Thermus thermophilus : Crystallization and X-ray analysis
Van Nerom, Katleen ; Tamman, Hedvig ; Takada, Hiraku ; Hauryliuk, Vasili LU
and Garcia-Pino, Abel
(2019)
In Acta Crystallographica Section F: Structural Biology Communications
p.561-569
- Abstract
The stringent response, controlled by (p)ppGpp, enables bacteria to trigger a strong phenotypic resetting that is crucial to cope with adverse environmental changes and is required for stress survival and virulence. In the bacterial cell, (p)ppGpp levels are regulated by the concerted opposing activities of RSH (RelA/SpoT homologue) enzymes that can transfer a pyrophosphate group of ATP to the 3′ position of GDP (or GTP) or remove the 3′ pyrophosphate moiety from (p)ppGpp. Bifunctional Rel enzymes are notoriously difficult to crystallize owing to poor stability and a propensity for aggregation, usually leading to a loss of biological activity after purification. Here, the production, biochemical analysis and crystallization of the... (More)
The stringent response, controlled by (p)ppGpp, enables bacteria to trigger a strong phenotypic resetting that is crucial to cope with adverse environmental changes and is required for stress survival and virulence. In the bacterial cell, (p)ppGpp levels are regulated by the concerted opposing activities of RSH (RelA/SpoT homologue) enzymes that can transfer a pyrophosphate group of ATP to the 3′ position of GDP (or GTP) or remove the 3′ pyrophosphate moiety from (p)ppGpp. Bifunctional Rel enzymes are notoriously difficult to crystallize owing to poor stability and a propensity for aggregation, usually leading to a loss of biological activity after purification. Here, the production, biochemical analysis and crystallization of the bifunctional catalytic region of the Rel stringent factor from Thermus thermophilus (Rel Tt NTD) in the resting state and bound to nucleotides are described. Rel Tt and Rel Tt NTD are monomers in solution that are stabilized by the binding of Mn2+ and mellitic acid. Rel Tt NTD crystallizes in space group P4122, with unit-cell parameters a = b = 88.4, c = 182.7 Å, at 4°C and in space group P41212, with unit-cell parameters a = b = 105.7, c = 241.4 Å, at 20°C.
(Less)
- author
- Van Nerom, Katleen
; Tamman, Hedvig
; Takada, Hiraku
; Hauryliuk, Vasili
LU
and Garcia-Pino, Abel
- publishing date
- 2019
- type
- Contribution to journal
- publication status
- published
- keywords
- Bacterial alarmone, PppGpp, Rel/RelA/SpoT, Stringent response, Thermus thermophilus
- in
- Acta Crystallographica Section F: Structural Biology Communications
- pages
- 561 - 569
- publisher
- Wiley-Blackwell
- external identifiers
-
- scopus:85070531465
- pmid:31397328
- ISSN
- 2053-230X
- DOI
- 10.1107/S2053230X19010628
- language
- English
- LU publication?
- no
- additional info
- Publisher Copyright: © 2019 International Union of Crystallography. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.
- id
- 9a905b14-c9ae-4147-9b48-27a63b231992
- date added to LUP
- 2021-09-24 20:34:10
- date last changed
- 2024-04-20 11:58:19
@article{9a905b14-c9ae-4147-9b48-27a63b231992,
abstract = {{<p>The stringent response, controlled by (p)ppGpp, enables bacteria to trigger a strong phenotypic resetting that is crucial to cope with adverse environmental changes and is required for stress survival and virulence. In the bacterial cell, (p)ppGpp levels are regulated by the concerted opposing activities of RSH (RelA/SpoT homologue) enzymes that can transfer a pyrophosphate group of ATP to the 3′ position of GDP (or GTP) or remove the 3′ pyrophosphate moiety from (p)ppGpp. Bifunctional Rel enzymes are notoriously difficult to crystallize owing to poor stability and a propensity for aggregation, usually leading to a loss of biological activity after purification. Here, the production, biochemical analysis and crystallization of the bifunctional catalytic region of the Rel stringent factor from Thermus thermophilus (Rel Tt <sup>NTD</sup>) in the resting state and bound to nucleotides are described. Rel Tt and Rel Tt <sup>NTD</sup> are monomers in solution that are stabilized by the binding of Mn<sup>2+</sup> and mellitic acid. Rel Tt <sup>NTD</sup> crystallizes in space group P4122, with unit-cell parameters a = b = 88.4, c = 182.7 Å, at 4°C and in space group P41212, with unit-cell parameters a = b = 105.7, c = 241.4 Å, at 20°C.</p>}},
author = {{Van Nerom, Katleen and Tamman, Hedvig and Takada, Hiraku and Hauryliuk, Vasili and Garcia-Pino, Abel}},
issn = {{2053-230X}},
keywords = {{Bacterial alarmone; PppGpp; Rel/RelA/SpoT; Stringent response; Thermus thermophilus}},
language = {{eng}},
pages = {{561--569}},
publisher = {{Wiley-Blackwell}},
series = {{Acta Crystallographica Section F: Structural Biology Communications}},
title = {{The Rel stringent factor from Thermus thermophilus : Crystallization and X-ray analysis}},
url = {{http://dx.doi.org/10.1107/S2053230X19010628}},
doi = {{10.1107/S2053230X19010628}},
year = {{2019}},
}