title: The crystal structure and biochemical properties of DHBPS from Streptococcus pneumoniae, a potential anti-infective target for Gram-positive bacteria.
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.
authors: Key Laboratory of Molecular Biology on Infectious Disease, Chongqing Medical University, YiXueYuanlu-1, Chongqing 400016, People's Republic of China; Department of Laboratory Medicine, Chongqing Medical University, YiXueYuanlu-1, Chongqing 400016, People's Republic of China.
authors: Jin
authors: J
authors: Li
authors: Zhou
authors: Z
authors: Hua
authors: Luo
authors: L
authors: Miao
authors: Tang
authors: T
authors: Jian
authors: Yang
authors: Y
authors: Wei
authors: Zhao
authors: Z
authors: Shasha
authors: Zhang
authors: Z
authors: Shaocheng
authors: Guo
authors: G
authors: Zhen
authors: Zhang
authors: Z
authors: Hongpeng
authors: Huang
authors: H
authors: Ailong
authors: Wang
authors: W
authors: Deqiang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D020134:Catalytic Domain; D016503:Drug Delivery Systems; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019751:Intramolecular Transferases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013296:Streptococcus pneumoniae / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D019751:Intramolecular Transferases; C066493:L-3,4-dihydroxy-2-butanone-4-phosphate synthase
keywords: 3,4-Dihydroxy-2-butanone-4-phosphate synthase; 3,4-dihydroxy-2-butanone-4-phosphate; 3,4-dihydroxy-2-butanone-4-phosphate synthase; 3,4-dihydroxy-2-butanone-4-phosphate synthase of Streptococcus pneumonia; Crystal structure; DHBP; DHBPS; DMSO; Enzyme analysis; PCR; PDB; Ru5P; S. pneumonia; SpDHBPS; Streptococcus pneumonia; Streptococcus pneumoniae; dimethyl sulfoxide; polymerase chain reaction; protein data bank; ribulose-5-phosphate