Number of the records: 1
Nonhydrolysable Analogues of (p)ppGpp and (p)ppApp Alarmone Nucleotides as Novel Molecular Tools
- 1.
SYSNO ASEP 0545506 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Nonhydrolysable Analogues of (p)ppGpp and (p)ppApp Alarmone Nucleotides as Novel Molecular Tools Author(s) Mojr, Viktor (UOCHB-X) ORCID
Roghanian, M. (SE)
Tamman, H. (BE)
Do Pham, Duy Dinh (UOCHB-X) ORCID
Petrová, Magdalena (UOCHB-X) RID
Pohl, Radek (UOCHB-X) RID, ORCID
Takada, H. (JP)
Van Nerom, K. (BE)
Ainelo, H. (BE)
Caballero-Montes, J. (BE)
Jimmy, S. (SE)
Garcia-Pino, A. (BE)
Hauryliuk, V. (EE)
Rejman, Dominik (UOCHB-X) RID, ORCIDSource Title ACS Chemical Biology. - : American Chemical Society - ISSN 1554-8929
Roč. 16, č. 9 (2021), s. 1680-1691Number of pages 12 s. Language eng - English Country US - United States Keywords pppGpp ; pppApp ; nonhydrolysable ; molecular tools OECD category Organic chemistry R&D Projects 8F19006 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Limited access Institutional support UOCHB-X - RVO:61388963 UT WOS 000697343400008 DOI 10.1021/acschembio.1c00398 Annotation While alarmone nucleotides guanosine-3′,5′-bisdiphosphate (ppGpp) and guanosine-5′-triphosphate-3′-diphosphate (pppGpp) are archetypical bacterial second messengers, their adenosine analogues ppApp (adenosine-3′,5′-bisdiphosphate) and pppApp (adenosine-5′-triphosphate-3′-diphosphate) are toxic effectors that abrogate bacterial growth. The alarmones are both synthesized and degraded by the members of the RelA-SpoT Homologue (RSH) enzyme family. Because of the chemical and enzymatic liability of (p)ppGpp and (p)ppApp, these alarmones are prone to degradation during structural biology experiments. To overcome this limitation, we have established an efficient and straightforward procedure for synthesizing nonhydrolysable (p)ppNuNpp analogues starting from 3′-azido-3′-deoxyribonucleotides as key intermediates. To demonstrate the utility of (p)ppGNpp as a molecular tool, we show that (i) as an HD substrate mimic, ppGNpp competes with ppGpp to inhibit the enzymatic activity of human MESH1 Small Alarmone Hyrolase, SAH, and (ii) mimicking the allosteric effects of (p)ppGpp, (p)ppGNpp acts as a positive regulator of the synthetase activity of long ribosome-associated RSHs Rel and RelA. Finally, by solving the structure of the N-terminal domain region (NTD) of T. thermophilus Rel complexed with pppGNpp, we show that as an HD substrate mimic, the analogue serves as a bona fide orthosteric regulator that promotes the same intra-NTD structural rearrangements as the native substrate. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2022 Electronic address https://doi.org/10.1021/acschembio.1c00398
Number of the records: 1