Screening and identification of novel cGAS homologues using a combination of in vitro and in vivo protein synthesis

Simple item page
dc.contributor.authorRolf, Jascha
dc.contributor.authorSiedentop, Regine
dc.contributor.authorLütz, Stephan
dc.contributor.authorRosenthal, Katrin
dc.date.accessioned2020-01-21T13:01:59Z
dc.date.available2020-01-21T13:01:59Z
dc.date.issued2019-12-22
dc.description.abstractThe cyclic GMP-AMP synthase (cGAS) catalyzes the synthesis of the multifunctional second messenger, cGAMP, in metazoans. Although numerous cGAS homologues are predicted in protein databases, the catalytic activity towards cGAMP synthesis has been proven for only four of them. Therefore, we selected five novel and yet uncharacterized cGAS homologues, which cover a broad range in the field of vertebrates. Cell-free protein synthesis (CFPS) was used for a pre-screening to investigate if the cGAS genes originating from higher organisms can be efficiently expressed in a bacterial expression system. As all tested cGAS variants were expressible, enzymes were synthesized in vivo to supply higher amounts for a subsequent in vitro activity assay. The assays were carried out with purified enzymes and revealed vast differences in the activity of the homologues. For the first time, the cGAS homologues from the Przewalski’s horse, naked mole-rat, bald eagle, and zebrafish were proven to catalyze the synthesis of cGAMP. The extension of the list of described cGAS variants enables the acquisition of further knowledge about the structural and molecular mechanism of cGAS, potentially leading to functional improvement of the enzyme.en
dc.identifier.urihttp://hdl.handle.net/2003/38533
dc.identifier.urihttp://dx.doi.org/10.17877/DE290R-20452
dc.language.isoende
dc.relation.ispartofseriesInternational Journal of Molecular Sciences;2020, 21(1), 105
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectcGASen
dc.subjectNucleotidyltransferaseen
dc.subjectcGAMPen
dc.subjectSTINGen
dc.subjectEnzyme screeningen
dc.subjectCFPSen
dc.subjectTXTLen
dc.subjectHeterologous proteinsen
dc.subject.ddc660
dc.titleScreening and identification of novel cGAS homologues using a combination of in vitro and in vivo protein synthesisen
dc.typeTextde
dc.type.publicationtypearticlede
dcterms.accessRightsopen access
eldorado.secondarypublicationtruede
eldorado.secondarypublication.primarycitationInternational Journal of Molecular Sciences. 2020, 21(1), S. 105de
eldorado.secondarypublication.primaryidentifierhttps://doi.org/10.3390/ijms21010105de

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
ijms-21-00105.pdf
Size:
2.7 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
4.85 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

Arbeitsgruppe Bioverfahrenstechnik