|Detection of Human Phospho-LRP‑6 (S1490) by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line transfected with human LRP-6 untreated (-) or treated (+) with 100 ng/mL Recombinant Human Wnt‑3a (High Purity) (Catalog # 5036-WNP) for 15 minutes. PVDF Membrane was probed with 1 µg/mL of Rabbit Anti-Human Phospho-LRP‑6 (S1490) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6649) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-LRP‑6 (S1490) at approximately 200 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 300 U/mL CIP for 1 hour. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
LRP6 (Low-density lipoprotein receptor-related protein 6) is a 200-210 kDa member of the LDLR family of proteins. It is widely expressed, and serves as a coreceptor for both Wnt and parathyroid hormone. In the Wnt system, LRP6 associates with select Fzd multipass receptors; in the PTH system, LRP6 complexes with PTHR1. Mature human LRP6 is a 1594 amino acid (aa) type I transmembrane glycoprotein. It contains a 1351 aa extracellular region (aa 20-1370) plus a 220 aa cytoplasmic domain (aa 1394-1613). The cytoplasmic domain contains two palmitoylation sites, one ubiquitination residue, and multiple phosphorylation motifs. Ser1490 is a key residue that impacts multiple activities. It undergoes both constitutive and receptor activation-induced phosphorylation. At least three enzymes likely act at this site, including PKA (associated with PTHR1), GSK-3 (associated with Fzd) and Cyclin Y/PFTK1 (associated with G2/M of the cell cycle). In general, Ser1490 phosphorylation results in beta -catenin stabilization, followed by either gene transcription or beta -catenin binding to centrosomes.
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