PXD028599

PXD028599 is an original dataset announced via ProteomeXchange.

Title	Quantitative proteomics analysis of affinity-puQuantitative proteomics analysis of affinity-purified Tribolium castaneum JHR complexesrified Tribolium castaneum JHR complexes
Description	Juvenile 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 quantitative phosphoproteomcis analysis uncovered multiple phosphorylation sites in the TcMET protein, some of which were induced by methoprene. A functional bipartite nuclear localization signal, straddled by phosphorylated residues, was found within the disordered C-terminal region of TcMET. Our present characterization of the recombinant JHR is a primary step towards understanding JHR regulation.
HostingRepository	PRIDE
AnnounceDate	2022-02-16
AnnouncementXML	Submission_2022-02-16_10:06:37.236.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Thomas Nebl
SpeciesList	scientific name: Tribolium castaneum; NCBI TaxID: 7070;
ModificationList	phosphorylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2021-09-16 23:37:25	ID requested
⏵ 1	2022-02-16 10:06:38	announced

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]

submitter keyword: Tribolium castaneum, Juvenile Hormone Receptor, LC-MSMS, DME labelling, Quantitative MS

Dr Thomas Nebl
contact affiliation	CSIRO Biomedical Manufacturing Biology R&D Group
contact email	tom.nebl@csiro.au
lab head
Thomas Nebl
contact affiliation	CSIRO
contact email	tom.nebl@csiro.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD028599
     1. Label: PRIDE project
     2. Name: Quantitative proteomics analysis of affinity-puQuantitative proteomics analysis of affinity-purified Tribolium castaneum JHR complexesrified Tribolium castaneum JHR complexes