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Published on: June 21, 2021
Enabling Global Analysis of Protein Citrullination via Biotin Thiol Tag-Assisted Mass Spectrometry
Yatao Shi1, Zihui Li2, Bin Wang1
1School of Pharmacy, University of Wisconsin─Madison, Madison, Wisconsin 53705, United States.
Abstract:
Citrullination is a key post-translational modification (PTM) that affects protein structures and functions. Although it has been linked to various biological processes and disease pathogenesis, the underlying mechanism remains poorly understood due to a lack of effective tools to enrich, detect, and localize this PTM. Herein, we report the design and development of a biotin thiol tag that enables derivatization, enrichment, and confident identification of citrullination via mass spectrometry. We perform global mapping of the citrullination proteome of mouse tissues. In total, we identify 691 citrullination sites from 432 proteins which represents the largest data set to date. We discover novel distribution and functions of this PTM. This study depicts a landscape of protein citrullination and lays the foundation for further deciphering their physiological and pathological roles.
Insights
Researchers developed a new biotin thiol tag to identify protein citrullination, a key modification in health and disease. This method identified the largest dataset of citrullination sites to date, revealing new functions and roles.
Area of Science:
- Biochemistry
- Proteomics
- Molecular Biology
Background:
- Protein citrullination is a critical post-translational modification (PTM) impacting protein structure and function.
- Understanding citrullination's role in biological processes and disease pathogenesis is limited by a lack of effective detection tools.
Purpose of the Study:
- To develop and validate a novel biotin thiol tag for the enrichment and identification of citrullination.
- To perform a global proteome-wide mapping of citrullination sites in mouse tissues.
Main Methods:
- Design and synthesis of a biotin thiol tag for citrullination derivatization.
- Application of the tag for enrichment and mass spectrometry-based identification of citrullinated proteins.
- Global citrullination profiling of mouse tissue proteomes.
Main Results:
- Successful development of a biotin thiol tag enabling confident citrullination identification.
- Identification of 691 citrullination sites across 432 proteins, establishing the largest dataset to date.
- Discovery of novel distributions and functions associated with protein citrullination.
Conclusions:
- The developed biotin thiol tag is an effective tool for studying protein citrullination.
- This study provides a comprehensive landscape of protein citrullination, crucial for understanding its physiological and pathological significance.
- The findings lay the groundwork for future research into the roles of citrullination in various biological contexts.
