Article Title: Mutation in the TRKB cholesterol recognition site that blocks antidepressant binding does not influence the basal or BDNF-stimulated activation of TRKB
Figure Lengend Snippet: (A) The sequence of TRKB transmembrane (TM) domain from rat (UniProt: Q63604, residues 430-453) was mutated at Y433 residue (Y433F) or the entire motif was substituted by the rat sequence of TRKA TM (P35739, residues 419-442). (B) The interaction between biotinylated BDNF (bBDNF) and TRKB is not affected by the TRKB.Y433F mutation or in TRKB/TRKA.TM constructs. (C) TRKB.Y433F does not prevent the BDNF-induced dimerization of TRKB. The constructs used allow the formation of TRKB.wt homodimer or TRKB.wt/Y433F heterodimer. (D , E) Analysis of TRKB phosphorylation shows that, cortical cultures from TRKB.Y433F heterozygous mouse embryos respond to BDNF similarly to the the TRKB.wt littermates, regardless of the tyrosine residue tested ( D : Y515, E : Y816).
Article Snippet: The plates were then washed 3x with PBS buffer, and various concentrations of biotinylated BDNF (Alomone Labs, cat#B-250, 0.1 – 100 pM) was added for 1h at RT.
Techniques: Sequencing, Mutagenesis, Construct