General assay for enzymes in the heptose biosynthesis pathways using electrospray ionization mass spectrometry

Jimin Park, Daeun Lee, Eun Kyoung Seo, Jae Sang Ryu, Dong Hae Shin

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

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.

Original languageEnglish
Pages (from-to)4521-4532
Number of pages12
JournalApplied Microbiology and Biotechnology
Volume101
Issue number11
DOIs
StatePublished - 1 Jun 2017

Keywords

  • Burkholderia pseudomallei
  • Electrospray ionization mass spectroscopy
  • Heptose biosynthesis pathways
  • Melioidosis
  • Nucleotide-activated heptose

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