FIGURE 2.

IDR-1 binds to the ZZ domain of p62. A, schematic representation of the constructs used for mapping of the p62 domains that interact with IDR-1. SH2, an SH2 binding domain; AID, an acidic interaction domain (PB1 domain comprises the SH2 and AID); LB, LIM protein Ajuba binding domain; PEST, amino acids Pro, Glu, Ser, Thr-enriched domain; LIR, LC3-interacting region; UBA, ubiquitin-associated domain. B, analysis of IDR-1 pulldown assays with constructs presented in A. Lysates from HEK293T cells overexpressing FLAG-tagged deletion mutants of human p62 were pulled down against biocytin (B) or biotinylated IDR-1 (P). Cell lysates (L) were used as control. Proteins pulled down were immunoblotted with anti-FLAG antibody.