Btk regulation in human and mouse B cells via protein kinase C phosphorylation of IBtk {gamma

The inhibitor of Bruton tyrosine kinase (IBtk) is a negative regulator of the Bruton tyrosine kinase (Btk), which plays a major role in B-cell differentiation; however, the mechanisms of IBtk-mediated regulation of Btk are unknown. Here we report that B-cell receptor (BCR) triggering caused serine-phosphorylation of IBtk at protein kinase C consensus sites and dissociation from Btk. By liquid chromatography and mass-mass spectrometry and functional analysis, we identified IBtk-S87 and -S90 as the critical amino acid residues that regulate the IBtk binding affinity to Btk. Consistently, the mutants IBtk carrying S87A and S90A mutations bound constitutively to Btk and down-regulated Ca2 fluxes and NF-B activation on BCR triggering. Accordingly, spleen B cells from Ibtk/ mice showed an increased activation of Btk, as evaluated by Y551-phosphorylation and sustained Ca2 mobilization on BCR engagement. These findings identify a novel pathway of Btk regulation via protein kinase C phosphorylation of IBtk. (Blood. 2011;117(24):6520-6531)