PXD070617

PXD070617 is an original dataset announced via ProteomeXchange.

Title	Human enteric α-defensin 5 is essential for intestinal barrier integrity by promoting focal adhesions via P2Y11-Rac1 activation
Description	Besides its well-documented antimicrobial functions, human enteric α-defensins 5 (HD5) has been recently found to act as a "double-edged sword" in intestinal defense by enhancing Shigella infection through inducing filopodial-like extensions via activating P2Y11 receptor in human colonic epithelial cells. Nonetheless, the physiological implications of HD5-P2Y11 interaction, beyond its involvement in Shigella infection, have not been comprehensively elaborated. Here, by using primary human colonic epithelial cells, human colonoid organoids, and a gut-on-a-chip model, we showed that HD5 plays an essential roles in establishing and restoring the human intestinal barrier by promoting cell adhesion in suspension or detached cells. Mechanistic investigation demonstrates that although P2Y11 still serves as the responsible receptor mediating the the barrier-protecting effect of HD5, a cell state-dependent alternative downstream signaling cascade was uncovered. In well-adhered cells, HD5-P2Y11 signaling preferentially engages the cAMP-PKA pathway to induce exploratory protrusions like filopodia. Conversely, in suspension cells or detached cells where adhesion is compromised, the signaling shifts priority to the FAK pathway to rapidly reinforce cell adhesion. This dynamic switching allows HD5 to precisely orchestrate epithelial barrier maintenance and repair. This pathway involves the phosphorylation and activation of FAK at key tyrosine residues (Tyr397, Tyr576, Tyr925), which in turn activates the small GTPase Rac1. This HD5-P2Y11-FAK-Rac1 axis enhances barrier function by promoting the formation and stabilization of focal adhesions, accelerating cell attachment, and facilitating epithelial migration for efficient repair. By delineating this novel pathway and its context-dependent regulation, the study fills the knowledge gap to understand the paradoxical roles of HD5 in human intestinal physiology and more importantly, provides a new perspective to understand the physiological significance of this human-specific defensin, extending its role beyond direct antimicrobial action to a key regulator of cytoskeletal dynamics and intestinal homeostasis.
HostingRepository	iProX
AnnounceDate	2025-11-12
AnnouncementXML	Submission_2025-11-11_19:04:44.776.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Mengyao Guo
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2025-11-11 19:03:27	ID requested
⏵ 1	2025-11-11 19:04:45	announced

Dataset with its publication pending

submitter keyword: human defensin 5, P2Y11, cell adhesion, focal adhesion

Dan Xu
contact affiliation	Xi'an Jiaotong University
contact email	dan.xu@xjtu.edu.cn
lab head
Mengyao Guo
contact affiliation	Xi'an Jiaotong University
contact email	myguo123@stu.xjtu.edu.cn
dataset submitter

iProX dataset URI