PXD017192

PXD017192 is an original dataset announced via ProteomeXchange.

Title	Epidermal Growth Factor-Induced Protein Phosphorylation Changes in Rat Inner Medullary Collecting Duct
Description	Epidermal growth factor (EGF) is a potent mitogenic agent promoting cell differentiation, proliferation, and growth. The kidney thick ascending limb of Henle’s loop and distal convoluted tubule were identified as major sites of EGF synthesis. As expected, EGF can modulate the functioning of kidney collecting duct including osmotic water permeability (Pf). Here, we found that EGF (0.1 uM) reduced the vasopressin-stimulated Pf by 26% in isolated perfused rat inner medullary collecting duct (IMCD). Immunoblotting using IMCD suspensions prepared in similar condition showed that EGF significantly reduced phosphorylation of AQP2 at Ser264 and Ser269 but had not affected on Ser256 or proline-directed Ser261 site. Quantitative phosphoproteomics was carried out to investigate other effects of EGF. Rat IMCD suspensions were treated with 1 M of EGF or vehicle for 30 min. Samples from three replicates of experiment were analyzed by TMT isobaric labeling quantitative protein mass spectrometry. We identified 23859 phosphopeptides with unique phosphorylation sites in EGF-treated rat IMCD samples, with 25 proteins increased and 100 proteins decreased in phosphorylation as determined by dual statistical analysis. EGF induced phosphorylation of multiple residues at the C-terminus of EGF receptor, EGFR or Erbb1, and mildly activated the classical MAPK pathway, RAF-MEK-ERK. EGF also affected phosphorylation of several proteins in PI3K/Akt pathway involved in cap-dependent translation initiation (Eif4ebp1, Gigyf2, Eif3b, Eif4g3, Pdcd4) and translation elongation (Eef2k, Eef2), predicted an overall translation repression. In comparison with a dDAVP-induced phosphoproteome, we found that the majority of proteins had phosphorylation change on specific site by either EGF or dDAVP but not both, suggesting a highly selectivity of phosphorylation modification induced by these two agents. Among the 25 peptides that had phosphorylation change on the same site by both EGF and dDAVP, AQP2 at Ser264/Ser269 was changed in opposite direction. We concluded that the short-term effect of EGF is insufficient to cause an effect on Ser261 phosphorylation of AQP2 in normal rat IMCD.
HostingRepository	PRIDE
AnnounceDate	2025-06-26
AnnouncementXML	Submission_2025-06-26_10:54:16.738.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Chung-Lin Chou
SpeciesList	scientific name: Rattus norvegicus (Rat); NCBI TaxID: 10116;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; phosphorylated residue; monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2020-01-20 02:56:41	ID requested
⏵ 1	2025-06-26 10:54:17	announced

Chou CL, Jayatissa NU, Kichula ET, Ou SM, Limbutara K, Knepper MA, Phosphoproteomic response to epidermal growth factor in native rat inner medullary collecting duct. Am J Physiol Renal Physiol, 328(1):F29-F47(2025) [pubmed]

10.1152/ajprenal.00182.2024;

submitter keyword: water, mass spectrometry,vasopressin, kidney, MAPK, protein kinase, ERBB

Mark Knepper
contact affiliation	Epithelial Systems Biology Laboratory National Heart, Lung, and Blood Institute National Institutes of Health
contact email	knepperm@nhlbi.nih.gov
lab head
Chung-Lin Chou
contact affiliation	National Institutes of Health
contact email	chouj@nhlbi.nih.gov
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD017192
     1. Label: PRIDE project
     2. Name: Epidermal Growth Factor-Induced Protein Phosphorylation Changes in Rat Inner Medullary Collecting Duct