A pectic polysaccharide isolated from the roots of Bupleurum falcatum L. stimulates the tyrosine phosphorylation of lipid rafts of murine B cells.

Bupleuran 2IIc, a pectic polysaccharide isolated from the roots of Bupleurum falcatum L., was previously characterized as a T cell-independent B cell mitogen. The endo-(1-->4)-alpha-D-polygalacturonase-resistant moiety of bupleuran 2IIc (bupleuran 2IIc/PG-1) was the active site for expression of the activity, and expression of the cyclin D2 gene by bupleuran 2IIc/PG-1 may be mediated via activation of Src family tyrosine kinase, phosphatidylinositol 3-kinase (PI 3-K) and phospholipase C (PLC)-gamma followed by activation of protein kinase C (PKC) and calcium mobilization (Matsumoto et al., Int. Immunopharmacol., 5, 1373-1386 (2005)). Plasma membrane microdomains (lipid rafts) are enriched in signaling molecules and suggested to be involved in numerous cell functions, including membrane traffic and signaling. When B cells were stimulated with bupleuran 2IIc/PG-1, clustering of membrane lipid rafts was observed. To consider whether lipid rafts are implicated in bupleuran 2IIc/PG-1-mediated B cell proliferation, we analyzed the phosphorylation of tyrosine residues of proteins in lipid rafts. When murine B cells were stimulated with bupleuran 2IIc/PG-1, tyrosine phosphorylation of proteins in lipid rafts fraction was observed within 5 min. Tyrosine phosphorylation in lipid rafts fraction by bupleuran 2IIc/PG-1 was inhibited by the Src-family tyrosine kinase inhibitor, PP2. Together with previously published data, the results presented in this study suggest that activation of signaling molecules in lipid rafts by stimulation of bupleuran 2IIc/PG-1 contributes to B cell proliferation as the membrane-proximal signaling event.