Brain quantitative proteomics combining GeLC-MS and isotope-coded protein labeling (ICPL)

Area of Science:

• Biochemistry
• Analytical Chemistry
• Neuroscience

Background:

• Mass spectrometry advancements have transformed proteomics.
• Quantitative proteomic methods, including in vitro stable isotope labeling, have concurrently evolved.
• Isotope-coded protein labeling (ICPL) is a key technique for proteome quantification.

Purpose of the Study:

• To present a detailed protocol for using ICPL within a shotgun proteomics workflow.
• To outline the advantages and disadvantages of the ICPL method.
• To demonstrate the applicability of ICPL for studying proteomes, with a focus on brain disorders.

Main Methods:

• The protocol involves specific sample preparation for ICPL.
• Protein identification is a prerequisite for the workflow.
• Shotgun proteomics techniques are employed for comprehensive analysis.

Main Results:

• The presented protocol is adaptable to various proteomes.
• ICPL provides quantitative insights into proteomic changes.
• The method's utility is exemplified in the context of brain disorder research.

Conclusions:

• ICPL is a valuable tool for quantitative proteomics.
• The protocol facilitates the study of complex proteomes, including those relevant to neurological conditions.
• Understanding the advantages and disadvantages of ICPL is crucial for its effective implementation.