Comprehensive Proteomic Analysis of Lipid Rafts from RBL-2H3 Mast Cells

Mast cells have long been recognized to have a direct and critical role in allergic and inflammatory reactions. In allergic diseases, these cells exert both local and systemic responses, including allergic rhinitis and anaphylaxis. Mast cell mediators are also related to many chronic inflammatory conditions. Besides the roles in pathological conditions, the biological functions of mast cells include roles in innate immunity, involvement in host defense mechanisms against parasites, immunomodulation of the immune system, tissue repair, and angiogenesis. Despite their growing significance in physiological and pathological conditions, much still remains to be learned about mast cell biology. This paper presents evidence that lipid rafts or raft components modulate many of the biological processes in mast cells, such as degranulation and endocytosis, play a role in mast cell development and recruitment, and contribute to the overall preservation of mast cell structure and organization.

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Proteomic Analysis of Lipid Rafts from RBL-2H3 Mast Cells

International Journal of Molecular Sciences ◽

10.3390/ijms20163904 ◽

2019 ◽

Vol 20 (16) ◽

pp. 3904 ◽

Cited By ~ 2

Author(s):

Edismauro Garcia Freitas Filho ◽

Luiz Augusto Marin Jaca ◽

Lilian Cristiane Baeza ◽

Célia Maria de Almeida Soares ◽

Clayton Luiz Borges ◽

...

Keyword(s):

Mast Cells ◽

Lipid Rafts ◽

Lipid Raft ◽

Protein Composition ◽

Enrichment Analysis ◽

Functional Enrichment Analysis ◽

Cell Types ◽

Cellular Membrane ◽

Functional Enrichment ◽

Membrane Microdomains

Lipid rafts are highly ordered membrane microdomains enriched in cholesterol, glycosphingolipids, and certain proteins. They are involved in the regulation of cellular processes in diverse cell types, including mast cells (MCs). The MC lipid raft protein composition was assessed using qualitative mass spectrometric characterization of the proteome from detergent-resistant membrane fractions from RBL-2H3 MCs. Using two different post-isolation treatment methods, a total of 949 lipid raft associated proteins were identified. The majority of these MC lipid raft proteins had already been described in the RaftProtV2 database and are among highest cited/experimentally validated lipid raft proteins. Additionally, more than half of the identified proteins had lipid modifications and/or transmembrane domains. Classification of identified proteins into functional categories showed that the proteins were associated with cellular membrane compartments, and with some biological and molecular functions, such as regulation, localization, binding, catalytic activity, and response to stimulus. Furthermore, functional enrichment analysis demonstrated an intimate involvement of identified proteins with various aspects of MC biological processes, especially those related to regulated secretion, organization/stabilization of macromolecules complexes, and signal transduction. This study represents the first comprehensive proteomic profile of MC lipid rafts and provides additional information to elucidate immunoregulatory functions coordinated by raft proteins in MCs.

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