PXD020321

PXD020321 is an original dataset announced via ProteomeXchange.

Title	Quantitative analysis of design substrates
Description	Proteases are crucial physiologic regulators of protein structure and function. While proteomic methods contributed extensively to protease characterization efforts, technical challenges remain in terms of throughput, scalability and large datasets of protease cleavages remain scarce. Here, we describe a high-throughput protease screen (HTPS) which allows simultaneous characterization of multiple proteases under various conditions on a microscale 96FASP format. After benchmarking the performance with Trypsin, LysC, GluC, AspN, Chymotrypsin, MMP-2 and MMP-3, we applied it to profile proteases of the blood coagulation cascade (Factors VIIa, IXa, Xa, XIa, Thrombin α, β and γ, Plasmin and Protein C). The large dataset enabled us to map activity, specificity, cleave entropy, allosteric changes of blood cascade proteases induced by Na+ and to predict potential substrates in a data-driven way. Here, we designed two octapeptides, GIPRAAGD (α Thrombin) and GIGRRIAE (Factor Xa), and evaluated their cleavage with the target proteases.
HostingRepository	PRIDE
AnnounceDate	2021-02-04
AnnouncementXML	Submission_2021-02-04_11:15:04.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Federico Uliana
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2020-07-13 00:19:35	ID requested
⏵ 1	2021-02-04 11:15:05	announced

Dataset with its publication pending

submitter keyword: blood coagulation proteases, specificity, entropy, allostery, substrates, high-throughput protease screen

Ruedi Aebersold
contact affiliation	Department of Biology, Institute of Molecular Systems Biology, ETH Zürich, Zürich, Switzerland Faculty of Science, University of Zürich, Zürich, Switzerland
contact email	aebersold@imsb.biol.ethz.ch
lab head
Federico Uliana
contact affiliation	ETH
contact email	uliana@imsb.biol.ethz.ch
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/2021/02/PXD020321

PRIDE project URI

• PRIDE
  1. PXD020321
     1. Label: PRIDE project
     2. Name: Quantitative analysis of design substrates