Metallo-β-lactamase (MBL) superfamily proteins have a common αβ/βα sandwich fold and perform a variety of functions through metal-mediated catalysis. However, because of the enormous scale of this superfamily, only a small percentage of the proteins belonging to the superfamily have been annotated structurally or functionally to date. Therefore, much remains unknown about the MBL superfamily proteins. Here, TW9814, a hypothetical MBL superfamily protein, was structurally and functionally investigated. Guided by the crystal structure of dimeric TW9814, it was demonstrated that TW9814 functions as a phosphodiesterase (PDE) in the presence of divalent metal ions such as manganese(II) or nickel(II). A docking model between TW9814 and the substrate bis(p-nitrophenyl)phosphate (bpNPP) showed the importance of the dimerization of TW9814 for its bpNPP-hydrolyzing activity and for the interaction between the enzyme and the substrate. TW9814 showed outstanding catalytic efficiency (k cat/K m) under alkaline conditions compared with other PDEs. The activity of TW9814 appears to be regulated through a disulfide bond, which is a feature that is not present in other MBL superfamily members. This study provides a platform for the functional characterization of other hypothetical proteins of the MBL or other superfamilies.
|Number of pages||10|
|Journal||Acta Crystallographica Section D: Structural Biology|
|Publication status||Published - 2022 Apr 1|
Bibliographical noteFunding Information:
The following funding is acknowledged: National Research Foundation of Korea (grant No. 2020M3A9G7103934 to Weontae Lee); Ministry of Oceans and Fisheries (grant No. 15250103 to Sun-Shin Cha; grant No. 20180430 to Sun-Shin Cha).
All Science Journal Classification (ASJC) codes
- Structural Biology