A direct spectropolarimetric assay of arabinose 5-phosphate isomerase.
Aldose-Ketose Isomerases
/ analysis
Bacterial Proteins
/ metabolism
Catalysis
Circular Dichroism
/ methods
Enzyme Assays
/ methods
Francisella tularensis
/ metabolism
Lipopolysaccharides
/ metabolism
Pentosephosphates
/ metabolism
Ribulosephosphates
/ analysis
Substrate Specificity
Sugar Acids
/ metabolism
Sugar Phosphates
/ metabolism
Arabinose 5-phosphate isomerase
Assay method
Circular dichroism
Francisella tularensis
KDO
KdsD
Spectropolarimeter
Journal
Analytical biochemistry
ISSN: 1096-0309
Titre abrégé: Anal Biochem
Pays: United States
ID NLM: 0370535
Informations de publication
Date de publication:
01 06 2021
01 06 2021
Historique:
received:
13
10
2020
revised:
20
12
2020
accepted:
18
01
2021
pubmed:
16
3
2021
medline:
13
8
2021
entrez:
15
3
2021
Statut:
ppublish
Résumé
Arabinose 5-phosphate isomerase (API) catalyzes the reversible isomerization of Ribulose 5-phosphate (Ru5P) to Arabinose 5-Phosphate (Ar5P) for the production of 3-deoxy-2-octulosonic acid 8-phosphate (KDO), a component of bacterial lipopolysaccharide (LPS) of gram-negative bacteria. API is an attractive target for therapeutic development against gram-negative bacterial pathogens. The current assay method of API activity utilizes a general reaction for keto sugar determination in a secondary, 3-h color development reaction with 25 N sulfuric acid which poses hazard to both personnel and instrumentation. We therefore aimed to develop a more user friendly assay of the enzyme. Since Ru5P absorbs in the UV region and contains at least 2 chiral centers, it can be expected to display circular dichroism (CD). A wavelength scan revealed indeed Ru5P displays a pronounced negative ellipticity of 30,560 mDeg M
Identifiants
pubmed: 33716126
pii: S0003-2697(21)00017-8
doi: 10.1016/j.ab.2021.114116
pii:
doi:
Substances chimiques
3-deoxyoctulosonate 8-phosphate
0
Bacterial Proteins
0
Lipopolysaccharides
0
Pentosephosphates
0
Ribulosephosphates
0
Sugar Acids
0
Sugar Phosphates
0
2-keto-3-deoxyoctonate
1069-03-0
arabinose 5-phosphate
13137-52-5
ribulose 5-phosphate
4151-19-3
Aldose-Ketose Isomerases
EC 5.3.1.-
arabinose-5-phosphate isomerase
EC 5.3.1.13
Types de publication
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
114116Informations de copyright
Copyright © 2021 Elsevier Inc. All rights reserved.