1. The crystal structure and biochemical properties of DHBPS from Streptococcus pneumoniae, a potential anti-infective target for Gram-positive bacteria.
- Author
-
Li J, Hua Z, Miao L, Jian T, Wei Y, Shasha Z, Shaocheng Z, Zhen G, Hongpeng Z, Ailong H, and Deqiang W
- Subjects
- Amino Acid Sequence, Bacterial Proteins genetics, Bacterial Proteins isolation & purification, Bacterial Proteins metabolism, Catalytic Domain, Drug Delivery Systems, Enzyme Stability, Escherichia coli genetics, Escherichia coli metabolism, Intramolecular Transferases genetics, Intramolecular Transferases isolation & purification, Intramolecular Transferases metabolism, Models, Molecular, Molecular Sequence Data, Protein Conformation, Protein Multimerization, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins metabolism, Sequence Alignment, Streptococcus pneumoniae genetics, Bacterial Proteins chemistry, Intramolecular Transferases chemistry, Streptococcus pneumoniae enzymology
- Abstract
The enzymes involved in riboflavin biosynthesis are considered to be potential anti-bacterial drug targets because these proteins are essential in bacterial pathogens but are absent in humans. 3,4-dihydroxy-2-butanone-4-phosphate synthase (DHBPS) is one of the key enzymes in the biosynthesis of riboflavin. DHBPS catalyzes the conversion of ribulose-5-phosphate (Ru5P) to 3,4-Dihydroxy-2-butanone-4-phosphate (DHBP) and formate. The purified SpDHBPS enzyme, in the presence of Mg(2+) ion, catalyzed the conversion of Ru5P to DHBP at a rate of 109nmolmin(-1)mg(-1) with an apparent Km value of 181μM at 37°C. Surprisingly, our experiments first revealed that DHBPS showed activity in the presence of the trivalent metal ion, Fe(3+). Furthermore, we determined the crystal structure of DHBPS from Gram-positive bacteria, Streptococcus pneumoniae, with 2.0Å resolution. The overall architecture of SpDHBPS was similar to its homologs, which comprise one β-sheet (five-stranded) and eight α-helices, adopting a three-layered α-β-α sandwich fold. Similar to the homologs, gel-filtration experiments verified that the enzyme was arranged as a dimer. Although the overall fold of DHBPS was similar, the significant structural differences between the species at the active site region may be utilized to develop antibacterial agents that are species-specific., (Crown Copyright © 2013. Published by Elsevier Inc. All rights reserved.)
- Published
- 2013
- Full Text
- View/download PDF