PXD028394

PXD028394 is an original dataset announced via ProteomeXchange.

Title	LC-MSMS analyses of purified, recombinant TcJHR 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 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.
HostingRepository	PRIDE
AnnounceDate	2022-02-16
AnnouncementXML	Submission_2022-02-16_08:02:49.944.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD028394
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported 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	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-09-09 08:35:13	ID requested
⏵ 1	2022-02-16 08:02:51	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

Tom Nebl
contact affiliation	CSIRO Biomedical Manufacturing Protein Mass Spectrometry Lab Parkville, VIC 3052, Australia
contact email	tom.nebl@csiro.au
lab head
Thomas Nebl
contact affiliation	CSIRO
contact email	tom.nebl@csiro.au
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2022/02/PXD028394

PRIDE project URI

• PRIDE
  1. PXD028394
     1. Label: PRIDE project
     2. Name: LC-MSMS analyses of purified, recombinant TcJHR complexes