Discoidin, CUB, and LCCL domain containing 2 (DCBLD2) is a neuropilin-like transmembrane scaffolding receptor with known and anticipated roles in vascular remodeling and neuronal positioning. DCBLD2 is also up-regulated in several cancers and can drive glioblastomas downstream of activated epidermal growth factor receptor. While a few studies have shown either a positive or negative role for DCBLD2 in regulating growth factor receptor signaling, little is known about the conserved signaling features of DCBLD family members that drive their molecular activities. We previously identified DCBLD2 tyrosine phosphorylation sites in intracellular YxxP motifs that are required for the phosphorylation-dependent binding of the signaling adaptors CRK and CRKL (CT10 regulator of kinase and CRK-like). These intracellular YxxP motifs are highly conserved across vertebrates and between DCBLD family members. Here, we demonstrate that, as for DCBLD2, DCBLD1 YxxP motifs are required for CRKL–SH2 (Src homology 2) binding. We report that Src family kinases (SFKs) and Abl differentially promote the interaction between the CRKL–SH2 domain and DCBLD1 and DCBLD2, and while SFKs and Abl each promote DCBLD1 and DCBLD2 binding to the CRKL–SH2 domain, the effect of Abl is more pronounced for DCBLD1. Using high-performance liquid chromatography coupled with tandem mass spectrometry, we quantified phosphorylation at several YxxP sites in DCBLD1 and DCBLD2, mapping site-specific preferences for SFKs and Abl. Together, these data provide a platform to decipher the signaling mechanisms by which these novel receptors drive their biological activities.
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Cover Image
Cover Image
An outer membrane barrel assembly machinery (BAM) is in the bacterial outer membrane (between the red and blue plates). The BAM machinery contains one outer membrane protein BamA (green) and four lipoproteins BamB (yellow), BamC (magnet), BamD (orange) and BamC (cyan), which is responsible for insertion and assembly unfolded outer membrane proteins into the outer membrane. Please see article by Dong et al, pages 3951–3961 for further details. Image kindly provided by Changjiang Dong.
Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2
Anna M. Schmoker, Jaye L. Weinert, Kyle J. Kellett, Hannah E. Johnson, Ryan M. Joy, Marion E. Weir, Alicia M. Ebert, Bryan A. Ballif; Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2. Biochem J 1 December 2017; 474 (23): 3963–3984. doi: https://doi.org/10.1042/BCJ20170615
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