Analysis of CD40 interactome in primary B cells

ABSTRACT: CD40 (also known as TNFRSF5) is a critical transmembrane molecule expressed on B cells and on myeloid cells including DCs. In B cells, CD40 engagement by its ligand CD40L (CD154) is essential to promote immunoglobulin class switching, memory differentiation, and germinal center formation. Using affinity purification coupled with mass spectrometry (AP-MS) analysis, we defined the composition and dynamics of the signaling protein complex used by CD40. To that aim, we used a knock-in mouse that expresses a chimeric CD40 molecule composed of a human extracellular domain, a human transmembrane segment, fused to a mouse intra-cytoplasmic tail in which a OneStregTag (OST) has been added (hCD40OST). We purified the protein complexes formed around CD40 using Streptactin beads, from B cells either non stimulated or stimulated for 30s, 120s, 300s or 600s by a monoclonal antibody that binds to human CD40 (Anti-CD40 Ab). Each AP-MS purification is associated with a corresponding control (purification from non-edited WT mouse B cells stimulated in the same conditions). The number of replicate biological experiments was n=3 for all time-points, and each sample was analyzed twice by single-run nano LC-MS.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Spleen, B Cell

SUBMITTER: Anne Gonzalez de Peredo  

LAB HEAD: Anne Gonzalez de Peredo

PROVIDER: PXD042695 | Pride | 2026-10-26

REPOSITORIES: Pride