PXD068305

PXD068305 is an original dataset announced via ProteomeXchange.

Title	Systematic mapping of the adipocyte plasma membrane proteome reveals KCC1 and PIT2 as novel insulin-responsive transporters
Description	The plasma membrane (PM) is a dynamic interface that integrates environmental cues with cellular responses. Insulin is known to remodel the PM primarily by stimulating the translocation of glucose transporter type 4 (GLUT4), but the full scope of this regulation remains poorly defined. Here, we performed a comprehensive analysis of insulin-regulated PM proteins in adipocytes by combining nine independent proteomic datasets generated using complementary PM enrichment strategies. This approach identified 66 high-confidence insulin regulated protein candidates, the majority (56 proteins) of which have not previously been implicated in insulin action. Among these, we experimentally validated the insulin-stimulated translocation of two transporters: potassium-chloride cotransporter 1 KCC1 (SLC12A4) and sodium-dependent phosphate transporter PIT2 (SLC20A2). Live-cell imaging and biochemical assays revealed that both proteins localize across multiple insulin-sensitive endosomal compartments, undergo dose-dependent trafficking to the PM, and require PI3K-AKT signaling for their mobilization. siRNA-mediated knockdown of KCC1 or PIT2 impaired insulin-stimulated glucose transport and their expression in adipose tissue correlated with human metabolic traits, suggesting a role for these transporters in insulin action. Strikingly, insulin-induced translocation of KCC1 and PIT2 was impaired in cells rendered insulin resistant by chronic hyperinsulinemia. These findings suggest that insulin resistance perturbs a broad set of endosomal trafficking proteins, likely impacting cellular functions beyond just glucose metabolism. Together, our work provides the first systematic map of insulin-regulated PM remodeling in adipocytes, establishes KCC1 and PIT2 as novel insulin-responsive transporters, and highlights new pathways through which insulin may coordinate ionic balance, phosphate homeostasis, and nutrient uptake.
HostingRepository	PRIDE
AnnounceDate	2026-03-23
AnnouncementXML	Submission_2026-03-22_17:37:22.123.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David James
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: NEWT:10090;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2025-09-12 00:23:13	ID requested
⏵ 1	2026-03-22 17:37:23	announced

10.1016/j.jbc.2026.111282;

Zhang Y, Cooke KC, Scavuzzo J, Cutler HB, Madsen S, Kearney AL, Conway OJ, Hawkins BL, Menon D, Humphrey SJ, Koumanov F, St, ö, ckli J, Geddes TA, Fazakerley DJ, Diaz-Vegas A, Burchfield JG, James DE, Integrated analysis of the adipocyte plasma membrane proteome reveals KCC1 and PIT2 as novel insulin-responsive transporters. J Biol Chem, 302(4):111282(2026) [pubmed]

submitter keyword: None

David James
contact affiliation	Charles Perkins Centre, The University of Sydney
contact email	david.james@sydney.edu.au
lab head
David James
contact affiliation	The University of Sydney
contact email	david.james@sydney.edu.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/2026/03/PXD068305

PRIDE project URI

• PRIDE
  1. PXD068305
     1. Label: PRIDE project
     2. Name: Systematic mapping of the adipocyte plasma membrane proteome reveals KCC1 and PIT2 as novel insulin-responsive transporters