PXD025907

PXD025907 is an original dataset announced via ProteomeXchange.

Title	Alternate covalent form of platelet αIIbβ3
Description	The αIIbβ3 integrin receptor coordinates platelet adhesion, activation and mechanosensing in thrombosis and haemostasis. Using differential cysteine alkylation and mass spectrometry, we have identified a disulfide bond in the αIIb subunit linking cysteines 490 and 545 that is missing in about one in three integrin molecules on the resting and activated human platelet surface. This alternate covalent form of αIIbβ3 is pre-determined as it is also produced by human megakaryoblasts and bovine hamster kidney cells transfected with recombinant human integrin. From co-immunoprecipitation experiments, the alternate form selectively partitions into focal adhesions on the activated platelet surface. Its function was evaluated in bovine hamster kidney cells expressing a mutant integrin with an ablated C490-C545 disulfide bond. The disulfide mutant integrin has extended residency time in focal adhesions due to reduced rate of clathrin-mediated integrin internalisation and recycling that is associated with enhanced affinity of the αIIb subunit for clathrin adaptor protein-2. The alternate covalent form also has different conformational dynamics measured by monoclonal antibody binding and molecular dynamic simulations, which is reflected in reduced fibrinogen binding and outside-in signalling. These findings indicate that the αIIbβ3 integrin receptor is produced in different covalent forms that have different functions. The C490, C545 cysteine pair is conserved across all 18 integrin α subunits and the disulfide bond in αV in a cancer cell line is similarly missing, suggesting that this alternate integrin form and function is also conserved.
HostingRepository	PRIDE
AnnounceDate	2022-02-17
AnnouncementXML	Submission_2022-02-17_03:15:54.929.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Aster Pijning
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2021-05-09 23:15:38	ID requested
⏵ 1	2022-02-17 03:15:56	announced

Pijning AE, Blyth MT, Coote ML, Passam F, Chiu J, Hogg PJ, 3 integrin that resides in focal adhesions and has altered function. Blood, 138(15):1359-1372(2021) [pubmed]

submitter keyword: Platelet, integrin, disulfide, covalent, αIIbβ3

Philip J. Hogg
contact affiliation	Chair in Translational Cancer Research, NHMRC Clinical Trials Centre, University of Sydney, Australia. NH&MRC Senior Principal Research Fellow. Deputy Director, Centenary Institute. Deputy Director, Sydney Catalyst. Head, ACRF Centenary Cancer Research Centre.
contact email	Phil.hogg@sydney.edu.au
lab head
Aster Pijning
contact affiliation	Centenary Insitute, The University of Sydney
contact email	a.pijning@centenary.org.au
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/2022/02/PXD025907

PRIDE project URI

• PRIDE
  1. PXD025907
     1. Label: PRIDE project
     2. Name: Alternate covalent form of platelet αIIbβ3