Universität zu Köln

Annibal, Andrea ORCID: 0000-0003-4311-5417, Ripa, Roberto, Ballhysa, Eugen, Latza, Christian, Hochhard, Nadine and Antebi, Adam (2021). Mass spectrometric characterization of cyclic dinucleotides (CDNs) in vivo. Anal. Bioanal. Chem., 413 (26). S. 6457 - 6469. HEIDELBERG: SPRINGER HEIDELBERG. ISSN 1618-2650

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Abstract

Cyclic dinucleotides (CDNs) are key secondary messenger molecules produced by cyclic dinucleotide synthases that trigger various cellular signaling cascades from bacteria to vertebrates. In mammals, cyclic GMP-AMP synthase (cGAS) has been shown to bind to intracellular DNA and catalyze the production of the dinucleotide 2 ' 3 ' cGAMP, which signals downstream effectors to regulate immune function, interferon signaling, and the antiviral response. Despite the importance of CDNs, sensitive and accurate methods to measure their levels in vivo are lacking. Here, we report a novel LC-MS/MS method to quantify CDNs in vivo. We characterized the mass spectrometric behavior of four different biologically relevant CDNs (c-di-AMP, c-di-GMP, 3 ' 3 ' cGAMP, 2 ' 3 ' cGAMP) and provided a means of visually representing fragmentation resulting from collision-induced dissociation at different energies using collision energy breakdown graphs. We then validated the method and quantified CDNs in two in vivo systems, the bacteria Escherichia coli OP50 and the killifish Nothobranchius furzeri. We found that optimization of LC-MS/MS parameters is crucial to sensitivity and accuracy. These technical advances should help illuminate physiological and pathological roles of these CDNs in in vivo settings. Graphical abstract

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Annibal, Andrea UNSPECIFIED orcid.org/0000-0003-4311-5417 UNSPECIFIED Ripa, Roberto UNSPECIFIED UNSPECIFIED UNSPECIFIED Ballhysa, Eugen UNSPECIFIED UNSPECIFIED UNSPECIFIED Latza, Christian UNSPECIFIED UNSPECIFIED UNSPECIFIED Hochhard, Nadine UNSPECIFIED UNSPECIFIED UNSPECIFIED Antebi, Adam UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-569888
DOI:	10.1007/s00216-021-03628-6
Journal or Publication Title:	Anal. Bioanal. Chem.
Volume:	413
Number:	26
Page Range:	S. 6457 - 6469
Date:	2021
Publisher:	SPRINGER HEIDELBERG
Place of Publication:	HEIDELBERG
ISSN:	1618-2650
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DNA SENSOR CGAS; GMP-AMP; STING PATHWAY; 2ND-MESSENGER; CELL; QUANTIFICATION Multiple languages Biochemical Research Methods; Chemistry, Analytical Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/56988