Extraction and detection of guanosine 5'-diphosphate-3'-diphosphate in amino acid starvation cells of Clavibacter michiganensis.
Clavibacter michiganensis
Extraction
Guanosine 5′-diphosphate-3′-diphosphate
LC-MS/MS
UHPLC
Journal
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
ISSN: 1678-4405
Titre abrégé: Braz J Microbiol
Pays: Brazil
ID NLM: 101095924
Informations de publication
Date de publication:
Sep 2021
Sep 2021
Historique:
received:
19
10
2020
accepted:
29
03
2021
pubmed:
11
4
2021
medline:
15
12
2021
entrez:
10
4
2021
Statut:
ppublish
Résumé
Guanosine 5'-diphosphate-3'-diphosphate (ppGpp) is a small molecule nucleotide alarmone that can accumulate under the amino acid starvation state and trigger the stringent response. This study reported the extraction of ppGpp from the Gram-positive bacteria Clavibacter michiganensis through methods using formic acid, lysozyme, or methanol. Following extraction, ppGpp was detected through ultra-high-performance liquid chromatography (UHPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The methanol method showed the highest extraction efficiency for ppGpp among the three methods tested. C. michiganensis cells in exponential growth phase was induced in amino acid starvation by serine hydroxamate (SHX) and used for ppGpp extraction and detection. When using the methanol extraction method, the results showed that ppGpp concentrations in SHX-treated samples were 15.645 nM, 17.656 nM, 20.372 nM, and 19.280 nM at 0 min, 15 min, 30 min and 1 h, respectively, when detected using LC-MS/MS. This is the first report on ppGpp extraction and detection in Clavibacter providing a new idea and approach for nucleotide detection and extraction in bacteria.
Identifiants
pubmed: 33837930
doi: 10.1007/s42770-021-00488-1
pii: 10.1007/s42770-021-00488-1
pmc: PMC8324763
doi:
Substances chimiques
Amino Acids
0
Diphosphates
0
Guanosine Tetraphosphate
33503-72-9
Methanol
Y4S76JWI15
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
1573-1580Informations de copyright
© 2021. Sociedade Brasileira de Microbiologia.
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