Post-Translational Modification Profiling-Functional Proteomics for the Analysis of Immune Regulation
Avital Eisenberg-Lerner1, Ifat Regev2, Yifat Merbl3
1Department of Immunology, The Weizmann Institute of Science, 234 Herzl Street, Rehovot, 7610001, Israel. Avital.eisenberg@weizmann.ac.il.
Methods in Molecular Biology (Clifton, N.J.)
|August 16, 2017
Summary
This study introduces PTM-profiling, a novel method using protein microarrays to quantitatively screen posttranslational modifications (PTMs) in immune cells. This technique aids in understanding PTMs
Area of Science:
- Biochemistry
- Immunology
- Proteomics
Background:
- Posttranslational modifications (PTMs) are crucial for regulating protein function, localization, and stability.
- Protein microarrays enable large-scale screening of PTMs, advancing target identification and regulatory role elucidation.
- The immune system is a key area for studying PTM perturbations due to their known roles in immune function and dysfunction.
Purpose of the Study:
- To present a novel PTM-profiling system combining functional extract-based assays with protein microarray screening.
- To highlight the advantages of this system for quantitative and unbiased PTM identification under specific cellular conditions.
- To discuss the significance of PTM profiling in addressing fundamental questions in immunology, particularly concerning the ubiquitin system.
Main Methods:
- Development of a PTM-profiling system integrating functional extract-based activity assays with protein microarrays.
- Preparation of functional cell extracts from differentiated or induced immune cell cultures.
- Screening of PTMs on protein arrays, including detailed protocols and guidelines for data analysis.
Main Results:
- The PTM-profiling system allows for quantitative and unbiased identification of protein modifications.
- The methodology is applicable to studying PTMs in various cellular contexts, with a focus on the immune system.
- Protocols for cell extract preparation and PTM screening on protein arrays are described.
Conclusions:
- PTM-profiling offers a powerful approach to quantitatively study protein modifications in biological systems.
- This method is particularly valuable for investigating the role of PTMs, including ubiquitination, in immune function and disease.
- The described protocols facilitate basic research in immunology by enabling detailed analysis of PTMs in immune cells.
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