Proteomics

Dataset Information

0

Mapping specificity, entropy, allosteric changes and substrates in blood coagulation proteases by a high-throughput protease screen

ABSTRACT: 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.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

SUBMITTER: Matej Vizovisek  

LAB HEAD: Ruedi Aebersold

PROVIDER: PXD018976 | Pride | 2021-02-05

REPOSITORIES: Pride

Json Xml
altmetric image

Publications

Mapping specificity, cleavage entropy, allosteric changes and substrates of blood proteases in a high-throughput screen.

Uliana Federico F   Vizovišek Matej M   Acquasaliente Laura L   Ciuffa Rodolfo R   Fossati Andrea A   Frommelt Fabian F   Goetze Sandra S   Wollscheid Bernd B   Gstaiger Matthias M   De Filippis Vincenzo V   Auf dem Keller Ulrich U   Aebersold Ruedi R  

Nature communications 20210316 1

Proteases are among the largest protein families and critical regulators of biochemical processes like apoptosis and blood coagulation. Knowledge of proteases has been expanded by the development of proteomic approaches, however, technology for multiplexed screening of proteases within native environments is currently lacking behind. Here we introduce a simple method to profile protease activity based on isolation of protease products from native lysates using a 96FASP filter, their analysis in  ...[more]

Similar Datasets

Proteomics HTPS screen of coagulation proteases at 20C with NaCl, ChCl, LiCl
2021-02-04 | PXD022959 | Pride
Proteomics Characterization of WN NS3 and AspN proteases
2021-02-04 | PXD022973 | Pride
Proteomics Multiple experiments: Analysis of the HTPS FT; Kinetics of Thrombin and Chymotrypsin; Effect of temperature on Thrombin and Chymotrypsin
2021-02-04 | PXD022972 | Pride
Clinical Oxygen deficiency of the human bowel
| 2533504 | ecrin-mdr-crc
Proteomics Targeted quantification of octapeptide substrate of FII and FX
2021-02-04 | PXD020319 | Pride
Proteomics Quantitative analysis of design substrates
2021-02-04 | PXD020321 | Pride
Proteomics Target quantification of octapeptide for Thrombin and FX
2021-02-04 | PXD020320 | Pride
Transcriptomics Comparison of gene expression in mouse placentas with different combination of MT1-MMP and MT2-MMP alleles
2010-03-09 | E-GEOD-18604 | biostudies-arrayexpress
Proteomics Degradomic identification of membrane type 1-matrix metalloproteinase (MT1-MMP/MMP14) as an ADAMTS9 and ADAMTS20 substrate
2023-07-20 | PXD036612 | Pride
Transcriptomics RNA-seq of Lon and FtsH conditional mutants of Mycoplasma pneumoniae under depleting and inducing conditions
2020-11-10 | E-MTAB-8537 | biostudies-arrayexpress