PXD035743

PXD035743 is an original dataset announced via ProteomeXchange.

Title	Identification of Cyclin D3-CDK4 disulfide heterodimer
Description	Oxidants induce cell cycle arrest to halt cell proliferation; however, little is known about the redox-regulated effector proteins that mediate these processes. Here, we report a novel kinase-inhibitory disulfide bond in cyclin D-CDK4 and investigate its role in cell proliferation and PH. Oxidative modifications of cyclin D-CDK4 were detected in human pulmonary arterial smooth muscle cells (HPASMCs) and human pulmonary arterial endothelial cells (HPAECs). Cysteine to alanine mutants were generated, and cell cycle experiments were employed to characterize the nature of this reversible intermolecular disulfide bond. The functional role of the disulfide was delineated using in vitro kinase activity assays, HPASMCs and knock-in cells. Finally, the cyclin D-CDK4 disulfide was assessed in vivo in the pulmonary arteries and isolated HPASMCs of PAH patients, and in three preclinical models of PH. Cyclin D-CDK4 forms an oxidant-induced heterodimeric disulfide dimer between C7/8 and C135, respectively. This reversible modification forms in cells in vitro and in pulmonary arteries in vivo to inhibit cyclin D-CDK4 kinase activity and decrease retinoblastoma protein (Rb) phosphorylation. Correspondingly, treatment of HPASMCs with H2O2 or auranofin induces cell cycle arrest. Notably, mutation of CDK4 C135 causes a kinase-impaired phenotype, which decreases the proliferation rate of cells, suggesting this cysteine is indispensable for cyclin D-CDK4 kinase activity. Pulmonary arteries and HPASMCs from patients with pulmonary arterial hypertension (PAH) display a decreased level of CDK4 disulfide, consistent with CDK4 being hyperactive in PAH. Furthermore, auranofin treatment, which induces the cyclin D-CDK4 disulfide, attenuates disease severity in experimental models of PH, by mitigating pulmonary vascular remodeling. A novel disulfide bond in cyclin D-CDK4 acts as a rapid switch to inhibit kinase activity and halt cell proliferation. This oxidative modification forms at a critical cysteine residue, which is unique to CDK4, offering the potential for design of a selective covalent inhibitor that may prove beneficial in pulmonary hypertension
HostingRepository	PRIDE
AnnounceDate	2026-01-16
AnnouncementXML	Submission_2026-01-16_02:43:24.952.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD035743
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Steven Lynham
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	dihydroxylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2022-08-02 10:21:17	ID requested
⏵ 1	2026-01-16 02:43:25	announced

10.1161/CIRCRESAHA.122.321836;

10.6019/PXD035743;

submitter keyword: CDK4, Disulfide, Cyclin D3, Oxidation

Dr Olena Rudyk
contact affiliation	Lecturer School of Cardiovascular and Metabolic Medicine & SciencesKing's College London
contact email	olena.rudyk@kcl.ac.uk
lab head
Steven Lynham
contact affiliation	King's College London
contact email	steven.lynham@kcl.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD035743
     1. Label: PRIDE project
     2. Name: Identification of Cyclin D3-CDK4 disulfide heterodimer