The Gram-negative bacterium Yersinia pseudotuberculosis is the causative agent of yersiniosis. D-glycero-α-D-manno-heptose-1-phosphate guanylyltransferase (HddC) is the fourth enzyme of the GDP-D-glycero-α-D-manno-heptose biosynthesis pathway which is important for the virulence of the microorganism. Therefore, HddC is a potential target of antibiotics against yersiniosis. In this study, HddC from the synthesized HddC gene of Y. pseudotuberculosis has been expressed, purified, crystallized. Synchrotron X-ray data from a selenomethionine-substituted HddC crystal were also collected and its structure was determined at 2.0 Å resolution. Structure analyses revealed that it belongs to the glycosyltransferase A type superfamily members with the signature motif GXGXR for nucleotide binding. Despite of remarkable structural similarity, HddC uses GTP for catalysis instead of CTP and UTP which are used for other major family members, cytidylyltransferase and uridylyltransferase, respectively. We suggest that EXXPLGTGGA and L(S/A/G)X(S/G) motifs are probably essential to bind with GTP and a FSFE motif with substrate.
|Number of pages||6|
|Journal||Biochimica et Biophysica Acta - Proteins and Proteomics|
|State||Published - Mar 2018|
Bibliographical noteFunding Information:
We are grateful to the staffs at both Pohang Light Source and Advanced Light Source. This work was supported by the Basic Science Research Program ( 2015R1D1A1A01058942 ) funded by the National Research Foundation of Korea grant granted by the Ministry of Education, Science and Technology , Republic of Korea (MEST). J. Park was supported by Brain Korea 21 (BK21) Project.
- D-glycero-α-D-manno-heptose-1-phosphate guanylyltransferase (HddC)
- GDP-D-glycero-α-D-manno-heptose biosynthesis pathway
- Glycosyltransferase a type superfamilyl
- X-ray crystal structure