Redox modificationomics is an emerging field of science that focuses on the study of the effects of protein redox modifications on cellular function. This research area combines techniques from chemistry, biology, and proteomics to provide new insights into intracellular signaling, metabolic regulation, and disease pathogenesis. The redox state of a protein directly affects its function and intracellular interactions. By studying redox modifications, scientists can gain a deeper understanding of how proteins respond to redox changes inside and outside cells, and how these changes affect cellular behavior and disease processes.
Figure 1Protein oxidation modification mass spectrometry.
In redox modification omics, mass spectrometry is widely used to identify and quantify redox modified proteins and sites. This technique relies on ionizing proteins or peptides and then identifying them by mass ratio by measuring the mass of those ions.
Process steps
1) Sample preparation: First, protein samples are processed by various methods (e.g., reduction, alkylation) to stabilize the redox state of the protein.
2) Enzymatic cleavage: Enzymatic digestion of proteins into smaller peptides for mass spectrometry analysis.
3) Mass spectrometry:
1.Ionization: Peptides are ionized in a mass spectrometer (e.g., by electrospray ionization [ESI] or matrix-assisted laser desorption ionization [MALDI]). 2.Mass analysis: Ionized peptides are massed by mass spectrometry (e.g., time-flight [TOF], ion trap, Fourier transform ion cyclotron resonance [FT-ICR]). 3.Tandem mass spectrometry: Selected ions are further broken down to obtain amino acid sequence information. 4) Identification and quantification:
1.Identification: Identify specific proteins and modification sites by comparing mass spectrometry data to databases. 2.Quantitation: Protein expression levels are quantified by measuring the abundance of specific peptides. Commonly used quantification methods include labeling (e.g., labeling) and non-labeling methods (e.g., labeling-free quantification). In redox modification omics, mass spectrometry also presents some special challenges:
1.Dynamics of modifications: Redox modifications are often reversible and dynamic, so these changes need to be captured quickly and accurately. 2.Diversity of modifications: Redox modifications may include various types such as sulfur oxidation, nitrosylation, etc., which requires mass spectrometry techniques to have sufficient resolution and sensitivity to distinguish and detect these modifications. Redox modificationomics is an interdisciplinary field that involves multiple disciplines such as biochemistry, molecular biology, and genetics. With the deepening of research, this field is expected to provide new insights into our understanding of complex biological processes and disease mechanisms.