Mast cell surfaceome characterization reveals CD98 heavy chain is critical for optimal cell function - 03/02/22
Abstract |
Background |
Mast cells are involved in many distinct pathologic conditions, suggesting that they recognize and respond to various stimuli and thus require a rich repertoire of cell surface proteins. However, mast cell surface proteomes have not been comprehensively characterized.
Objective |
We aimed to further characterize the mast cell surface proteome to obtain a better understanding of how mast cells function in health and disease.
Methods |
We enriched for glycosylated surface proteins expressed in mouse bone marrow–derived cultured mast cells (BMCMCs) and identified them using mass spectrometry analysis. The presence of novel surface proteins in mast cells was validated by real-time quantitative PCR and flow cytometry analysis in BMCMCs and peritoneal mast cells (PMCs). We developed a clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing approach to disrupt genes of interest in BMCMCs.
Results |
The glycoprotein enrichment approach resulted in the identification of 1270 proteins in BMCMCs, 378 of which were localized to the plasma membrane. The most common protein classes among plasma membrane proteins were small GTPases, receptors, and transporters. One such cell surface protein was CD98 heavy chain (CD98hc), encoded by the Slc3a2 gene. Slc3a2 gene disruption resulted in a significant reduction in CD98hc expression, adhesion, and proliferation.
Conclusions |
Glycoprotein enrichment coupled with mass spectrometry can be used to identify novel surface molecules in mast cells. Moreover, CD98hc plays an important role in mast cell function.
Le texte complet de cet article est disponible en PDF.Key words : Mast cells, glycoprotein enrichment, mass spectrometry, CRISPR/Cas9, surfaceome, CD98hc
Abbreviations used : BMCMC, Cas9, CD, CD98hc, CFSE, CRISPR, crRNA, ERK, FcεRIβ, FcεRIγ, FN, GO, ImmGen, LAMP1, LAT, mRNA, MS, PMC, RNP, SCF
Plan
| Supported by funding from the National Institutes of Health (grant R21 AI144231 to A.M.P.). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. |
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| Disclosure of potential conflict of interest: The authors declare that they have no relevant conflicts of interest. |
Vol 149 - N° 2
P. 685-697 - février 2022 Retour au numéro
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