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General assay for enzymes in the heptose biosynthesis pathways using electrospray ionization mass spectrometry.
- Source :
-
Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2017 Jun; Vol. 101 (11), pp. 4521-4532. Date of Electronic Publication: 2017 Mar 09. - Publication Year :
- 2017
-
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.
- Subjects :
- Burkholderia pseudomallei drug effects
Inhibitory Concentration 50
Lignans pharmacology
Lipopolysaccharides biosynthesis
Nucleotidyltransferases antagonists & inhibitors
Nucleotidyltransferases chemistry
Nucleotidyltransferases drug effects
Phenols pharmacology
Spectrometry, Mass, Electrospray Ionization methods
Substrate Specificity
Biosynthetic Pathways
Burkholderia pseudomallei enzymology
Enzyme Assays methods
Heptoses biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1432-0614
- Volume :
- 101
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Applied microbiology and biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 28280867
- Full Text :
- https://doi.org/10.1007/s00253-017-8148-1