Related Experiment Video
Updated: Jan 28, 2026
05:30
Author Spotlight: Advancing the Analysis of Plasma Extracellular Vesicle Proteome for Cardiovascular Biomarker Studies
Published on: January 31, 2025
1.1K
Proteomic discovery of protease substrates.
Oliver Schilling1, Christopher M Overall
1The UBC Centre for Blood Research, Departments of Oral Biological & Medical Sciences, and Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada.
Current Opinion in Chemical Biology
|December 30, 2006
Summary
Identifying protease substrates in vivo is challenging. New proteomic techniques, including mass spectrometry and SILAC pulse-chase, improve the study of protease activity and substrate identification in biological systems.
Area of Science:
- Biochemistry
- Proteomics
- Molecular Biology
Background:
- Identifying protease substrates in vivo is difficult due to the vast number of proteins and transient substrate levels.
- Proteomics has advanced significantly, offering sensitive methods for quantitative analysis.
Purpose of the Study:
- To review and highlight advancements in technologies for identifying protease substrates in vivo.
- To discuss the application of these technologies in understanding protease function and pathology.
Main Methods:
- Mass spectrometry-based global proteome analysis coupled with liquid chromatography and stable isotope labeling (e.g., ICAT, iTRAQ, SILAC).
- Quantitative analysis of proteolytically generated neo amino-termini (terminopes).
- Activity-based probes for functional protease profiling.
Main Results:
- These methods have increased coverage and sensitivity for quantitative proteomics.
- Numerous new matrix metalloproteinase substrates have been identified.
- In vivo SILAC pulse-chase experiments allow the study of protein turnover and proteome dynamics.
Conclusions:
- A suite of complementary technologies is now available to dissect the 'protease web'.
- These tools are crucial for understanding protease function and its dysregulation in disease states.
- Advancements facilitate the study of protease activity and substrate identification in complex biological systems.
Related Concept Videos
01:26Drug Discovery: Overview
11.4K
Drug discovery is a multifaceted process involving extensive screening, testing, and optimization of lead compounds to identify potential new drugs for therapeutic use. It combines several approaches, including screening large numbers of natural products, chemical modification of known active molecules, identification of new drug targets, and rational design based on biological mechanisms and drug-receptor structure. These approaches are carried out in both academic research laboratories and...
11.4K
01:33Proteomics
9.7K
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
9.7K
02:18The Proteasome
10.2K
Eukaryotic cells can degrade proteins through several pathways. One of the most important amongst these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
10.2K
02:54Protein Kinases and Phosphatases
15.1K
Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
15.1K
01:19Enzyme Kinetics
104.0K
Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
104.0K
02:34Enzymes
94.6K
Inside living organisms, enzymes act as catalysts for many biochemical reactions involved in cellular metabolism. The role of enzymes is to reduce the activation energies of biochemical reactions by forming complexes with its substrates. The lowering of activation energies favor an increase in the rates of biochemical reactions.
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
94.6K