Deciphering the pH-dependent oligomerization of aspartate semialdehyde dehydrogenase from Wolbachia endosymbiont of Brugia malayi: An in vitro and in silico approaches.

The enzyme aspartate semialdehyde dehydrogenase (ASDH) plays a pivotal role in the amino acid biosynthesis pathway, making it an attractive target for the development of new antimicrobial drugs due to its absence in humans. This study aims to investigate the presence of ASDH in the filarial parasite Wolbachia endosymbiont of Brugia malayi (WBm) using both in vitro and in silico approaches. The size exclusion chromatography (SEC) and Native-PAGE analysis demonstrate that WBm -ASDH undergoes pH-dependent oligomerization and dimerization. To gain a deeper understanding of this phenomenon, the modelled monomer and dimer structures were subjected to pH-dependent dynamics simulations in various conditions. The results reveal that residues Val240, Gln161, Thr159, Tyr160, and Trp316 form strong hydrogen bond contacts in the intersurface area to maintain the structure in the dimeric form. Furthermore, the binding of NADP+ induces conformational changes, leading to an open or closed conformation in the structure. Importantly, the binding of NADP+ does not disturb either the dimerization or oligomerization of the protein, a finding confirmed through both in vitro and in silico analysis. These findings shed light on the structural characteristics of WBm -ASDH and offer valuable insights for the development of new inhibitors specific to WBm , thereby contributing to the development of potential therapies for filarial parasitic infections. [Display omitted] • ASDH from the Wolbachia endosymbiont of Brugia malayi (WBm) exists as a dimer in the solution state. • Furthermore, WBm -ASDH exhibits pH-dependent higher-order oligomerization and dimerization. • Under alkaline conditions, WBm -ASDH forms a higher-order oligomer, while under acidic pH conditions, it exists as a dimer. • Protein dimerization is facilitated by network communication between active site intersubunits. [ABSTRACT FROM AUTHOR]