The structure of pseudoenzyme PDX1.2 and its complex with active enzyme PDX1.3
EMSL Project ID
50635
Abstract
Aim - Determine the high-resolution structure of the protein PDX1.2 alone and in complex with PDX1.3. PDX1.2 is an important pseudo-enzyme with regulatory roles in vitamin B6 biosynthesis. It directly interacts with its catalytic analog PDX1.3 to stabilize and improve vitamin B6 synthesis under heat and oxidative stress. While the structure of PDX1.3 has been solved by X-ray crystallography, PDX1.2 remains uncharacterized at the molecular level. We have already identified optimal freezing conditions and have a low-resolution single particle cryo-EM dataset. Our goal with the current proposal is to use PNCC instrumentation to collect sufficient images to build the first high-resolution 3D atomic model of PDX1.2. We want to compare and understand similarities and differences between analogous active enzymes PDX1.1 and PDX1.3 (both resolved by X-ray) and the PDX1.2 regulatory pseudo-enzyme and hetero-complexes of PDX1.2/PDX1.3. The current study will be critical to understand this protein-protein network at the molecular level and dissect the mechanism of vitamin B6 biosynthesis under normal and stress conditions.
Project Details
Start Date
2018-12-01
End Date
2019-05-20
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Novikova I.V., N. Sharma, T.H. Moser, R.L. Sontag, Y. Liu, M. Collazo, and D.D. Cascio, et al. 2018. "Protein Structural Biology Using Cell-Free Platform from Wheat Germ." Advanced Structural and Chemical Imaging 4. PNNL-SA-136527. doi:10.1186/s40679-018-0062-9