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PXD031169-2

PXD031169 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleEngineering and exploiting synthetic allostery of NanoLuc luciferase
DescriptionThe ability to endow constitutively active proteins with synthetic allostery is a central objective of Synthetic Biology, mostly focused on the creation of protein biosensors – artificial sensing proteins with easily quantifiable outputs. Here we hypothesize that circular permutation of proteins increases the probability of functional coupling of new N- and C- terminal sequences with the active center of the protein through increased local structural disorder. We test this by constructing a synthetically allosteric version of circular permutated NanoLuc luciferase, activated through ligand-induced intramolecular non-covalent cyclisation. The developed biosensors cover a range of emission wavelengths and display sensitivity as low as 50 pM and dynamic range as high as 16-fold and could quantify their cognate ligand in human fluids such as saliva, serum and lysed blood. We apply hydrogen exchange kinetic mass spectrometry to analyze time resolved structural changes in the developed biosensors and observed ligand-mediated folding of newly created termini. While a large study is required for determining how frequent synthetic allostery emerges following circular permutation and what types of protein folds are susceptible to it, this study provides motivation and justification for such efforts.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_05:31:10.790.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterJonathan Phillips
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationListNo PTMs are included in the dataset
InstrumentSynapt MS
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-01-21 13:23:35ID requested
12022-02-10 00:57:45announced
22024-10-22 05:31:11announced2024-10-22: Updated project metadata.
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: LC-MS, HDMSE, HDX, Biosenso, allostery
Contact List
Jonathan Phillips
contact affiliationLiving Systems Institute, University of Exeter, EX4 4QD, UK
contact emailjj.phillips@exeter.ac.uk
lab head
Jonathan Phillips
contact affiliationLiving Systems Institute, University of Exeter
contact emailjj.phillips@exeter.ac.uk
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
Repository Record List
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