Structural elements of antizymes 1 and 2 are required for proteasomal degradation of ornithine decarboxylase

The antizymes constitute a conserved gene family with at least three mammalian orthologs. As described previously, in a degradation system utilizing rabbit reticulocyte lysate, antizyme 1 (AZ1) accelerates proteasomal ornithine decarboxylase (ODC) degradation, but antizyme 2 (AZ2) does not. To examine the relationship between antizyme structure and function, we further characterized the properties of AZ1 and AZ2 and protein chimeras composed of elements of the two. AZ1 binds to ODC with about a 3-fold higher potency than AZ2, but this cannot account for their distinct degradative activities. The dissimilar degradative capacity of AZ1 and AZ2 is also observed using purified proteasomes. A series of reciprocal AZ1/ AZ2 chimeras was used to determine the sequence elements needed to direct ODC degradation. An element contained within amino acids 130-145 of AZ1 is essential for this function. Constructs in which amino acids 130-145 were exchanged between the antizymes confirmed the critical nature of this region. Within this region amino acids 131 and 145 proved responsible for the functional difference between the two forms of AZ.