PXD031508

PXD031508 is an original dataset announced via ProteomeXchange.

Title	Dynamics of integrin α5β1, fibronectin, and their complex
Description	Integrin α5β1 mediates cell adhesion to the extracellular matrix (ECM) by binding to fibronectin (Fn). Recognition of both a RGD motif in the 10th type-III Fn domain (Fn10) and the synergy site in the 9th type-III Fn domain (Fn9) provide selectivity of α5β1 for fibronectin. Here, we have compared synergy site mutations and Fn truncation mutations for their α5β1 binding affinities and stabilities and interrogated Fn and α5β1 binding using hydrogen deuterium exchange mass spectrometry (HDX-MS) to gain insights into binding sites and the effects of ligand binding on integrin conformational change. Our results suggest that the synergistic effect of Fn9 requires both specific residues and a folded domain and that some residues that were considered to contribute to synergy are required for stability. HDX changes in fibronectin are localized to a peptide containing contacting residues in two β-strands and an intervening loop in Fn9 and the RGD containing loop in Fn10, indicative of binding sites. Decreases in HDX in α5β1 were found in an α5 peptide that associates with the Fn9 domain and inferred from a decrease in β1 subunit α1 helix exchange relative to an RGD peptide ligand. However, the dominant effect of Fn binding was an increase in deuterium exchange in α5β1 that was distributed over six sites in β1 and one in α5 that undergo changes in conformation or solvent accessibility and one unexpected site located in a contact between the α5 thigh and β-propeller domains.
HostingRepository	PRIDE
AnnounceDate	2022-08-09
AnnouncementXML	Submission_2022-08-09_06:50:09.527.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	John R. Engen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2022-02-09 02:34:44	ID requested
⏵ 1	2022-08-09 06:50:10	announced

Su Y, Iacob RE, Li J, Engen JR, Springer TA, 1, fibronectin, and their complex reveal sites of interaction and conformational change. J Biol Chem, 298(9):102323(2022) [pubmed]

submitter keyword: Integrin, fibronectin, affinity, conformational change, thermal stability, hydrogen deuterium exchange

John R. Engen
contact affiliation	Department of Chemistry & Chemical Biology, Northeastern University
contact email	j.engen@northeastern.edu
lab head
John R. Engen
contact affiliation	Northeastern University
contact email	j.engen@northeastern.edu
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/2022/08/PXD031508

PRIDE project URI

• PRIDE
  1. PXD031508
     1. Label: PRIDE project
     2. Name: Dynamics of integrin α5β1, fibronectin, and their complex