PXD059156-1

PXD059156 is an original dataset announced via ProteomeXchange.

Title	Identification of high molecular weight protein/protein complexes in human cultured cells (HEK293T) LC-MS/MS
Description	Phosphoribosyl pyrophosphate synthetase (PRPS) is a highly conserved enzyme that conducts the chokepoint reaction of nucleotide biosynthesis by converting ribose-5-phosphate (R-5-P) to phosphoribosyl pyrophosphate (PRPP). Due to the presence of multiple isoforms and related PRPS-associated proteins in opisthokont species, the precise nature of the PRPS enzyme in cells and tissues remains uncertain. Using proteomics approaches and biochemistry, we demonstrate that these individual components assemble together to form a heterogeneous megadalton complex comprising of PRPS1, PRPS2, and two PRPS associated proteins – PRPSAP1 and PRPSAP2. To validate this and identify other protein/protein complexes that exists in a similar high molecular weight (HMW) range in human (HEK293T) cells, we performed shotgun proteomics on size exclusion chromatography fractions containing HMW proteins/protein complexes. This analysis identified a total of 262 unique proteins, which included ribosomal proteins and CAD confirming enrichment for HMW proteins. This dataset also revealed that among the eight cytosolic enzymes residing in HMW range, two were PRPS isozymes, indicating that the PRPS
HostingRepository	PRIDE
AnnounceDate	2025-05-13
AnnouncementXML	Submission_2025-05-13_13:45:38.477.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Tom Cunningham
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	methylthiolated residue; phosphorylated residue; deamidated residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2024-12-22 20:43:35	ID requested
⏵ 1	2025-05-13 13:45:39	announced

10.1101/2024.10.01.616059;

Karki BR, Macmillan AC, Vicente-Mu, ñ, oz S, Greis KD, Romick LE, Cunningham JT, Evolutionary origins and innovations sculpting the mammalian PRPS enzyme complex. bioRxiv, ():(2024) [pubmed]

submitter keyword: PRPS complex, High molecular weight proteins, protein complexes,Megadalton assemblies, LC-MS/MS

Dr. Tom Cunningham
contact affiliation	University of Cincinnati College of Medicine
contact email	cunnijn@ucmail.uc.edu
lab head
Tom Cunningham
contact affiliation	Department of Cancer Biology, University of Cincinnati College of Medicine
contact email	cunnijn@ucmail.uc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD059156
     1. Label: PRIDE project
     2. Name: Identification of high molecular weight protein/protein complexes in human cultured cells (HEK293T) LC-MS/MS