Anti-ASB1 (aa 172-221) polyclonal antibody

The growing demand of pharmaceutical industry for more effective drugs requires new molecules with promising medicinal activities. In the present work, a natural product anisaldehyde was treated with hydrazine and 3,5-dichloroaniline to synthesize their Schiff bases, ASB1 and ASB2, which were assessed for various bioactivities. ASB1 was synthesized by conventional reflux method while ASB2 was synthesized by reflux as well as by mechanochemical grinding method which gave higher yield. The bases were recrystalised, and their structures were elucidated based on XRD and spectroscopic studies. Hirshfeld surface analysis was also carried out. They showed considerable urease inhibitory activity, almost comparable with the standard thiourea. The activity of ASB1 was much higher than ASB2. Acetylcholinesterase inhibitory activity of ASB1 was also higher than that of ASB2. The antioxidant activities were determined using DPPH, ABTS radical scavenging and total antioxidant capacity (TAC) assays. The bases were very poor scavengers of DPPH radical. However, they showed considerable anti-radical activity against ABTS radical, ASB2 being more active than ASB1, while ASB1 showed higher TAC than ASB2. In conclusion, the bases appeared to have good drugability as inhibitors of urease and acetylcholinesterase enzymes. They can be easily synthesized for possible large-scale applications. The grinding method proved to be more efficient than the reflux method.

The ankyrin repeat and SOCS box (ASB) family is composed of 18 proteins from ASB1 to ASB18 and belongs to the suppressor of cytokine signaling (SOCS) box protein superfamily. ASB2 was recently shown to interact with a certain Cul-Rbx module to form an E3 ubiquitin (Ub) ligase complex, but the functional composition of the ASB-containing E3 Ub ligase complexes remains to be characterized. Here, we show that ASB proteins interact with Cul5-Rbx2 but neither Cul2 nor Rbx1 in cells. Mutational analysis revealed that the highly conserved amino acid sequences of the BC box and Cul5 box in the SOCS box of ASB proteins were essential for the interaction with Cul5-Rbx2. Although ASB proteins show slight divergences from the consensus sequences of the BC box and Cul5 box, all five tested ASB proteins bound to Cul5-Rbx2. Furthermore, all three tested ASB complexes containing Cul5-Rbx2 were found to have E3 Ub ligase activity. These findings suggest that the ASB family proteins interact with Cul5-Rbx2 to form E3 Ub ligases and play significant roles via a ubiquitination-mediated pathway. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.