Protease Activity Profiling Via Programmable Phage Display
Roman-Melendez, G.; Venkataraman, T.; Monaco, D.; Larman, H. B.
Show abstract
Endopeptidases catalyze the internal cleavage of proteins, playing pivotal roles in protein turnover, substrate maturation and the activation of signaling cascades. A broad range of biological functions in health and disease are controlled by proteases, yet assays to characterize their activities at proteomic scale do not yet exist. To address this unmet need, we have developed SEPARATE (Sensing EndoPeptidase Activity via Release and recapture using flAnking Tag Epitopes), which uses monovalent phage display of the entire human proteome at 90-aa peptide resolution. We demonstrate that SEPARATE is compatible with several human proteases from distinct catalytic classes, including Caspase-1, ADAM17, and Thrombin. Both well-characterized and newly identified substrates of these enzymes were detected in the assay. SEPARATE was used to discover a non-canonical Caspase-1 substrate, the E3 ubiquitin ligase HUWE1, a key mediator of apoptotic cell death. SEPARATE is a novel methodology to enable efficient, unbiased assessment of endopeptidase activity using a phage-displayed proteome.
Matching journals
The top 13 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Engineered Reversible Inhibition of SpyCatcher Reactivity Enables Rapid Generation of Bispecific Antibodies 94%
- A Versatile Soluble Siglec Scaffold for Sensitive and Quantitative Detection of Glycan Ligands 94%
- Identification of motif-based interactions between SARS-CoV-2 protein domains and human peptide ligands pinpoint antiviral targets. 94%
Similar papers in this journal
- Systematic profiling of peptide substrate specificity in N-terminal processing by methionine aminopeptidase using mRNA display and an unnatural methionine analogue 95%
- Peptide-antibody Fusions Engineered by Phage Display Exhibit Ultrapotent and Broad Neutralization of SARS-CoV-2 Variants 94%
- A Novel Regioselective Approach to Cyclize Phage-Displayed Peptides in Combination with Epitope-Directed Selection to Identify a Potent Neutralizing Macrocyclic Peptide for SARS-CoV-2 93%
Similar papers in this journal
- Deep profiling of protease substrate specificity enabled by dual random and scanned human proteome substrate phage libraries 97%
- DNA origami demonstrate the unique stimulatory power of single pMHCs as T-cell antigens 93%
- Directed Evolution of Genetically Encoded LYTACs for Cell-Mediated Delivery 93%
Similar papers in this journal
- Neutralizing antibodies targeting the SARS-CoV-2 receptor binding domain isolated from a naïve human antibody library 93%
- Phage display assisted discovery of a pH-dependent anti-alpha-cobratoxin antibody from a natural variable domain library 93%
- Screening de novo designed protein binders in unpurified lysate using flow induced dispersion analysis 93%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.