Thymine+Diphosphate-Glucose+4,6+dehydratase+(Bacillus+anthracis)

Enzyme Name: thymine diphosphate ( dTDP)-Glucose 4,6-dehydratase (rfbB) EC Number: 4.2.1.46 PDB ID: 6BI4 Organism: //Bacillus anthracis// AA Length: 354 Gene Length: 1065 Involved in three pathways: (works with a group of proteins) Ligands on the X-ray structure are sucrose, a sulfate ion, NAD, and a nickel (II) ion. Substrate is dTDP-d-glucose dTDP-Glucose 4,6-dehydratase is involved in a pathway that synthesizes rhamnose sugar, a polysaccharide that is incredibly important in cell wall development of Gram-positive bacteria. The role of rhamnose in synthesis of the cell wall was studied in streptococcus and found to comprise 50-60% of the cell wall thickness, and that bacteria that did not produce this sugar had severe growth and cell division abnormalities. This could have the same effect in //Bacillus anthracis.// Link to PDB page: [] dTDP-glucose 4,6-dehydratase is not similar to any human proteins or even any proteins belonging to any organism in Kingdom Animalia. There are only a few compounds currently under study as inhibitors of dTDP-glucose 4,6-dehydratase. This means it would be a good enzyme to study because it is not being overstudied as a potential target for inhibiting //Bacillus anthracis.// dTDP-glucose 4,6-dehydratase is highly expressed in the organism because it plays a large role in the synthesis of the cell wall of //Bacillus anthracis.// Based on experiments that have already been completed, dTDP-glucose 4,6-dehydratase can be expressed in E. Coli using a plasmid called rfbB. In the majority of past completed assays, the plasmid was created from scratch using PCR. This plasmid was then transformed into E. coli and expressed. Gene sequence: Atgaaattcctcattaccggcggagccggctttataggctccgcagttatccgccacatcatcaaccacactcaagacagcgtggtcaaccttgataagttgacctacgcaggcaatcttgaatctttggcttcggttgcagacaacccgcgttatacatttgaacaggcagatatttgcgaccgctctgcgcttgagtggattttcttgcaccatcaaccggatgccgttatgcacttggccgcggaaagccatgtcgaccgctcgatcgacgggccggcggcgtttattgaaaccaatatcgtcggcacttataccttgttggaggcagcacgggcatactggcaaaacatgccgtctgaaaaacaggcgtcgttccgttttcatcatatttctaccgacgaagtgtatggcgatttgcacggtaccgacgatttgtttaccgaaaccacgccttatgcgcccagcagcccttattccgcttccaaagcctccagcgaccatttagtaagggcatggcggcgcacttacggcttgccttgtttgattaccaattgttccaacaactacggcccttatcattttcctgaaaaactgattcctttgatgattctcaatgccatcggcggcagacccttgcccgtttacggcaacggtttgcagatacgcgattggctgtttgtcgaagatcatgcgcgggctttatatgccgtgatcacacaggggaaaataggggaaacctataatatcggcggccataacgaaaaaaccaatctcgaagtggtgcaaaccgtctgcaaactgctggaagaattgatgcccgaaaaacccgccggcgttcgagcctatgaagatttgatcacgtttgtcgaagaccgccccggccacgatgtgcgttatgccattgatgcggccaaaatcaaccacgagcttggctggaagcccgaagaaacttttgaatcgggtatccgtaaaaccgtagcatggtatttgaacaaccgcggctggtggcaacgtgtgctcgacggctcctaccggctcgaacgcttgggagtggcaaaatga Amino Acid Sequence: MKFLITGGAGFIGSAVIRHIINHTQDSVVNLDKLTYAGNLESLASVADNPRYTFEQADICDRSALEWIFLHHQPDAVMHLAAESHVDRSIDGPAAFIETNIVGTYTLLEAARAYWQNMPSEKQASFRFHHISTDEVYGDLHGTDDLFTETTPYAPSSPYSASKASSDHLVRAWRRTYGLPCLITNCSNNYGPYHFPEKLIPLMILNAIGGRPLPVYGNGLQIRDWLFVEDHARALYAVITQGKIGETYNIGGHNEKTNLEVVQTVCKLLEELMPEKPAGVRAYEDLITFVEDRPGHDVRYAIDAAKINHELGWKPEETFESGIRKTVAWYLNNRGWWQRVLDGSYRLERLGVAK
 * 1) Streptomycin biosynthesis
 * 2) Polyketide sugar unit biosynthesis
 * 3) Biosynthesis of vancomycin group antibiotics

Assay Information: Substrates are readily available for purchase from Sigma. Absorbance assay at 320 nm using a microplate spectrophotometer.