BioID2-based proximity labeling identifies DDIT4-interacting proteins in human coronary artery smooth muscle cells

ABSTRACT: DDIT4 (DNA damage-inducible transcript 4) is a stress-responsive protein that has been implicated in vascular smooth muscle cell (VSMC) phenotypic switching and vascular remodeling. However, the comprehensive protein interaction network of DDIT4 in VSMCs remains poorly characterized. In this study, we employed BioID2-based proximity labeling to identify DDIT4-interacting proteins in primary human coronary artery smooth muscle cells (HCASMCs). HCASMCs were transduced with adenoviruses encoding BioID2-NC (negative control) or BioID2-DDIT4 fusion protein for 36 hours, followed by supplementation with 50 μM biotin (Sigma, B4501) for 16 hours to label proximal proteins. Biotinylated proteins were enriched using streptavidin-coupled magnetic beads (Invitrogen, 65601), separated by SDS-PAGE, and subjected to in-gel tryptic digestion. The resulting peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) on an Orbitrap Fusion Lumos mass spectrometer. Protein identification was performed by searching the acquired spectra against the UniProt protein database. This dataset provides a resource for understanding the DDIT4 interactome in VSMCs and may offer insights into the molecular mechanisms underlying DDIT4-mediated VSMC phenotypic modulation.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: fengfeng li  

LAB HEAD: Zhi Zeng

PROVIDER: PXD083369 | Pride | 2026-10-05

REPOSITORIES: Pride