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PXD028394

PXD028394 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleLC-MSMS analyses of purified, recombinant TcJHR complexes
DescriptionJuvenile hormone (JH) serves vital roles in insect reproduction, development, and many aspects of physiology. JH primarily acts at the gene-regulatory level through an intracellular receptor. The JH receptor (JHR) is a ligand-activated complex of transcription factors of the bHLH-PAS family, consisting of the JH-binding protein Methoprene-tolerant (MET) and its partner Taiman (TAI). Initial studies have indicated significance of post-transcriptional modification (phosphorylation), subunit assembly, and nucleocytoplasmic transport of JHR in JH signaling. However, our knowledge of JHR regulation at the protein level remains rudimentary, partly due to the difficulty of obtaining functional JHR proteins in a purified form. Here we present successful fermentation-level purification of JHR complexes of MET and TAI proteins from two insect species, the beetle Tribolium castaneum and the mosquito Aedes aegypti. The recombinant JHR subunits from each species were co-expressed using a baculovirus system in an insect cell line and purified through affinity steps, yielding soluble proteins, capable of binding both the hormonal ligand (JH III) and DNA bearing cognate JH-response elements. The present shotgun LC-MSMS analyses identified at least ten potential phosphoserine and phosphothreonine residues located in the C-terminal disordered region of TcMET. A functional bipartite nuclear localization signal, straddled by some of these phosphorylated residues, was subsequently identified within the disordered C-terminal region of TcMET.
HostingRepositoryPRIDE
AnnounceDate2022-02-16
AnnouncementXMLSubmission_2022-02-16_08:02:49.944.xml
DigitalObjectIdentifierhttps://dx.doi.org/10.6019/PXD028394
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterThomas Nebl
SpeciesList scientific name: Tribolium castaneum; NCBI TaxID: 7070;
ModificationListphosphorylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentQ Exactive HF
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-09-09 08:35:13ID requested
12022-02-16 08:02:51announced
Publication List
Jindra M, McKinstry WJ, Nebl T, Bittova L, Ren B, Shaw J, Phan T, Lu L, Low JKK, Mackay JP, Sparrow LG, Lovrecz GO, Hill RJ, Purification of an insect juvenile hormone receptor complex enables insights into its post-translational phosphorylation. J Biol Chem, 297(6):101387(2021) [pubmed]
Keyword List
submitter keyword: Tribolium castaneum, Juvenile Hormone Receptor, LC-MSMS
Contact List
Tom Nebl
contact affiliationCSIRO Biomedical Manufacturing Protein Mass Spectrometry Lab Parkville, VIC 3052, Australia
contact emailtom.nebl@csiro.au
lab head
Thomas Nebl
contact affiliationCSIRO
contact emailtom.nebl@csiro.au
dataset submitter
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