PXD037724

PXD037724 is an original dataset announced via ProteomeXchange.

Title	Enhanced Access to the Human Phosphoproteome with Genetically Encoded Phosphothreonine
Description	Protein phosphorylation is a ubiquitous post-translational modification that governs signaling cascades and protein-protein interactions. Orthogonal translation systems repurpose evolutionarily divergent aminoacyl-tRNA synthetase and tRNA pairs for the co-translational insertion of a modified amino acid. Subsequent advancements over the last decade have enabled the insertion of phospho-amino acids, bypassing a priori knowledge requirements of upstream kinases for the study of phosphoproteins. Here we optimized a pThrOTS and corresponding E. coli strain for pThr protein production. We then produced a peptidome library containing ~57,000 known threonine/phosphothreonine phosphosites using oligonucleotide library synthesis. We were able to identify approximately ~20% of the peptides encoded by the pThr library, and ~44% of the peptides encoded by the Thr library with mass spectrometry. Robust, genetically encoded phosphothreonine revealed a new activation and inhibition mechanism for the kinase CHK2. Proteome-wide surveys of interactions between active CHK2 and our peptidome library identified novel substrates and motif elements. Finally, we developed a novel technique, Hi-P+, for directly linking kinase substrate discovery to phospho-binding domain recognition, unveiling multi-level interaction networks with phosphosite resolution. This new methodology enables kinase-specific, proteome-wide surveys of multiple phosphorylation-dependent protein-protein interactions. This dataset is for pThr peptide library
HostingRepository	PRIDE
AnnounceDate	2022-11-11
AnnouncementXML	Submission_2022-11-11_09:39:26.787.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	JackMoen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-10-26 01:00:12	ID requested
⏵ 1	2022-11-11 09:39:27	announced

Dataset with its publication pending

submitter keyword: synthetic biology, phosphorylation,Phosphoprotein, peptide library

JesseRinehart
contact affiliation	Department of Cellular & Molecular Physiology, Yale School of Medicine Systems Biology Institute, Yale University
contact email	jesse.rinerhart@yale.edu
lab head
JackMoen
contact affiliation	Yale University
contact email	jack.moen@yale.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD037724
     1. Label: PRIDE project
     2. Name: Enhanced Access to the Human Phosphoproteome with Genetically Encoded Phosphothreonine