title: General assay for enzymes in the heptose biosynthesis pathways using electrospray ionization mass spectrometry.
abstract: The ADP-L-glycero-β-D-manno-heptose and the GDP-6-deoxy-α-D-manno-heptose biosynthesis pathways play important roles in constructing lipopolysaccharide of Gram-negative bacteria. Blocking the pathways is lethal or increases antibiotic susceptibility to pathogens. Therefore, the enzymes involved in the pathways are novel antibiotic drug targets. Here, we designed an efficient method to assay the whole enzymes in the pathways using mass spectrometry and screened 148 compounds. One promising lead is (-)-nyasol targeting D-glycero-α-D-manno-heptose-1-phosphate guanylyltransferase (HddC) included in the GDP-6-deoxy-α-D-manno-heptose biosynthesis pathway from Burkholderia pseudomallei. The inhibitory activity of the lead compound against HddC has been confirmed by blocking the system transferring the guanosine monophosphate (GMP) moiety to α-D-glucose-1-phosphate. (-)-Nyasol exhibits the half maximal inhibitory concentration (IC50) value of 17.6 μM. A further study is going on using (-)-nyasol derivatives to find better leads with high affinity.
authors.affiliation: Department of Pharmacy, College of Pharmacy and Graduate School of Pharmaceutical Sciences, Global Top 5 Research Program, Ewha W. University, Seoul, 03760, Republic of Korea.
authors.forename: Jimin
authors.initials: J
authors.lastname: Park
authors.affiliation: Department of Pharmacy, College of Pharmacy and Graduate School of Pharmaceutical Sciences, Global Top 5 Research Program, Ewha W. University, Seoul, 03760, Republic of Korea.
authors.forename: Daeun
authors.initials: D
authors.lastname: Lee
authors.affiliation: Department of Pharmacy, College of Pharmacy and Graduate School of Pharmaceutical Sciences, Global Top 5 Research Program, Ewha W. University, Seoul, 03760, Republic of Korea.
authors.forename: Eun Kyoung
authors.initials: EK
authors.lastname: Seo
authors.affiliation: Department of Pharmacy, College of Pharmacy and Graduate School of Pharmaceutical Sciences, Global Top 5 Research Program, Ewha W. University, Seoul, 03760, Republic of Korea.
authors.forename: Jae-Sang
authors.initials: JS
authors.lastname: Ryu
authors.affiliation: Department of Pharmacy, College of Pharmacy and Graduate School of Pharmaceutical Sciences, Global Top 5 Research Program, Ewha W. University, Seoul, 03760, Republic of Korea. dhshin55@ewha.ac.kr.
authors.forename: Dong Hae
authors.initials: DH
authors.lastname: Shin
mesh_terms: D053898:Biosynthetic Pathways; D016957:Burkholderia pseudomallei / Q000187:drug effects / Q000201:enzymology*; D057075:Enzyme Assays / Q000379:methods*; D006539:Heptoses / Q000096:biosynthesis*; D020128:Inhibitory Concentration 50; D017705:Lignans / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000096:biosynthesis; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000187:drug effects; D010636:Phenols / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013379:Substrate Specificity
chemical_list: D006539:Heptoses; D017705:Lignans; D008070:Lipopolysaccharides; D010636:Phenols; C109393:nyasol; D009713:Nucleotidyltransferases; C020842:guanylyltransferase
keywords: Burkholderia pseudomallei; Electrospray ionization mass spectroscopy; Heptose biosynthesis pathways; Melioidosis; Nucleotide-activated heptose