|Detection of Human, Mouse, and Rat Mind Bomb 1/MIB1 by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line, mouse liver tissue, and rat liver tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat Mind Bomb 1/MIB1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7305) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Mind Bomb 1/MIB1 at approximately 115 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
MIB1 (Mind bomb 1; also DIP-1 and ZZANK2) is a 110-115 kDa E3 ubiquitin ligase. It is widely expressed in both embryo and adult, and appears to have a non-redundant role in Notch receptor signaling. Notch signaling is initiated when a transmembrane (TM) Notch ligand (Jagged or Delta) binds to Notch on an adjacent cell. This ligation induces cleavage of the Notch extracellular domain (ECD) with the creation of a Jagged/Delta:NotchECD complex. This complex is now internalized by the Notch ligand-expressing cell. The displacement of the NotchECD from its TM segment promotes further Notch processing, resulting in Notch signaling. The key internalization step by the ligand-expressing cell is mediated by the MIB1 E3 ubiquitin ligase. Human MIB1 is 1006 amino acids (aa) in length. It contains multiple interspersed Ankyrin repeats, MIB domains, and zinc-finger motifs. There is one potential alternative start site at Met84. Over aa 365-461, human MIB1 shares 99% aa sequence identity with mouse MIB1.
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