Synapse Associated Protein 102 (SAP102) Binds the C-Terminal Part of the Scaffolding Protein Neurobeachin
Figure 1
Nbea interacts with a fraction of SAP102 in vivo.
(A) Schematic representation of mouse Nbea (NCBI Reference Sequence: NP_085098.1). The predicted armadillo (ARM) repeat-flanked Concavanalin A (Con A)-like lectin domain is localized at the N-terminus of the protein (blue). The region the Nbea antibody was raised against is depicted by the purple rectangle, encompassing also the PKA binding site (pink stripe). At the C-terminus the domain of unknown function 1088 (DUF; in orange), the Pleckstrin-Homology like domain (PH; in gray), the BEACH domain (yellow) and the WD40 repeats (red) are depicted. (B) Co-immunoprecipitation of SAP102 and Nbea from crude membrane with microsomes fraction (P2+M) of P84 WT mice. Proteins were immunoprecipitated (IP) with two different α-SAP102 antibodies, i.e. a mouse monoclonal (NeuroMab clone N19/2; left lane) and a rabbit polyclonal one (GenScript; right lane), respectively. In the control condition non-coated, empty beads (EB) were used for the IP. The Input lane represents the crude membrane with microsomes fraction that was used for immunoprecipitation. Immuno-blotting (IB) was performed with α-Nbea and α-SAP102 antibody (NeuroMab clone N19/2). (C) Dendritic Nbea immunoreactivity shows limited overlap with SAP102. DIV14 WT hippocampal neurons (E18) fixed in methanol and stained for endogenous SAP102 (in green), endogenous Nbea (in red) and MAP2 (not shown in the merge). Top scale bar = 20 µm, lower scale bar = 5 µm.
