Targeting bacterial membranes: NMR spectroscopy characterization of substrate recognition and binding requirements of D-arabinose-5-phosphate isomerase.
Lipopolysaccharide (LPS) is an essential component of the outer membrane of gram-negative bacteria and consists of three elements: lipid A, the core oligosaccharide, and the O-antigen. The inner-core region is highly conserved and contains at least one residue of 3-deoxy-D-manno-octulosonate (Kdo). Arabinose-5-phosphate isomerase (API) is an aldo-keto isomerase catalyzing the reversible isomerization of D-ribulose-5-phosphate (Ru5P) to D-arabinose-5-phosphate (A5P), the first step of Kdo biosynthesis. By exploiting saturation transfer difference (STD) NMR spectroscopy, the structural requirements necessary for API substrate recognition and binding were identified, with the aim of designing new API inhibitors. In addition, simple experimental conditions for the STD experiments to perform a fast, robust, and efficient screening of small libraries of potential API inhibitors, allowing the identification of new potential leads, were set up. Due to the essential role of API enzymes in LPS biosynthesis and gram-negative bacteria survival, by exploiting these data, a new generation of potent antibacterial drugs could be developed.
Airoldi
Cristina
C
Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, 20126, Milan, Italy.
Sommaruga
Silvia
S
Merlo
Silvia
S
Sperandeo
Paola
P
Cipolla
Laura
L
Polissi
Alessandra
A
Nicotra
Francesco
F
D019747:Aldose-Ketose Isomerases / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D004926:Escherichia coli / Q000378:metabolism; D007536:Isomerism; D008070:Lipopolysaccharides / Q000096:biosynthesis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008566:Membranes / Q000737:chemistry* / Q000378:metabolism; D019081:O Antigens / Q000096:biosynthesis*; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis; D013379:Substrate Specificity; D013400:Sugar Acids / Q000378:metabolism
D008070:Lipopolysaccharides; D019081:O Antigens; D011135:Polysaccharides, Bacterial; D013400:Sugar Acids; D019747:Aldose-Ketose Isomerases; C041486:arabinose-5-phosphate isomerase; C019714:D-arabinose isomerase