Fig. 1: DJ1 is endogenous substrate of CRL4CRBN.
From: CRBN modulates synuclein fibrillation via degradation of DNAJB1 in mouse model of Parkinson disease
a CRBN levels are high in the brains of AppNL-G-F mice, an AD mouse model as compared to wild type (WT) mice at the ages of 3 months (3 m) and 18 months (18 m). data was extracted from https://alzmap.org/ using “CRBN” as query. TH,thalamus; HYP,hippocampus; CNU,cerebralnucleus; CTXsp,corticalsubplate; OLF,olfactoryarea; ENTI,entorhinalarea; AUD,auditoryarea; SSp,primarysomatosensory; PLT,posteriorparietalassociationarea; RSP,retrosplenialarea. b Expression of certain chaperones (Heat shock proteins or HSP) and cochaperones (DNAJ proteins) is increased in the whole brain lysates of Crbn-/- (KO) mice as compared to Crbn+/+ (WT) mice. DJ2, DJ1, and DJ6 are marked in blue, coral-red, and green, respectively. c DJ1 is structural homolog of DJ2. d β-turn- β loop of DJ2 involved in binding to CRBN is structurally conserved in DJ1. e CRBN binds to DJ1. SH-SY5Y cells were transiently co-transfected with HA-CRBN and Myc-DJ1* shows non-specific band, ** shows heavy chain band. Cells were lysed, immunoprecipitated with IgG control or α-HA antibody. n = 5 (f) DJ1 undergoes CRBN-mediated ubiquitination. SH-SY5Y cells were transiently transfected with HA-Ub and lysed in Ubn buffer, IP was performed with α-DJ1 antibody, n = 5. g CRBN binds to DJ1 in the brain lysates of mouse. Endogenous CRBN was pulled down with α-CRBN antibody and immunoblotting was performed with the antibodies indicated, AMPK-α was used as positive control.
