Structure of fructose bisphosphate aldolase fromBartonella henselaebound to fructose 1,6-bisphosphate

Abstract

PDB Reference: fructose bisphosphate aldolase, 3mmt.Fructose bisphosphate aldolase (FBPA) enzymes have been found in a broad range of eukaryotic and prokaryotic organisms. FBPA catalyses the cleavage of fructose 1,6-bisphosphate into glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The SSGCID has reported several FBPA structures from pathogenic sources, including the bacterium Brucella melitensis and the protozoan Babesia bovis. Bioinformatic analysis of the Bartonella henselae genome revealed an FBPA homolog. The B. henselae FBPA enzyme was recombinantly expressed and purified for X-ray crystallographic studies. The purified enzyme crystallized in the apo form but failed to diffract; however, well diffracting crystals could be obtained by cocrystallization in the presence of the native substrate fructose 1,6-bisphosphate. A data set to 2.35 Å resolution was collected from a single crystal at 100 K. The crystal belonged to the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 72.39, b = 127.71, c = 157.63 Å . The structure was refined to a final free R factor of 22.2%. The structure shares the typical barrel tertiary structure and tetrameric quaternary structure reported for previous FBPA structures and exhibits the same Schiff base in the active site.

Structural and Functional Characterization of Methicillin-ResistantStaphylococcus aureus’sClass IIb Fructose 1,6-Bisphosphate Aldolase

“…Unlike the active site loop within class II FBAs, the β6α6 loop containing residues 177–191, and previously termed the Z-loop, is almost always well-defined structurally. ,,,,,,,,− This loop forms part of the substrate pocket as well as contains a histidine residue involved in coordination with the active site Zn­(II) ion . Between class IIb FBAs, such as HpFBA, whose structures have been reported, and SaFBA, HpFBA possess two additional residues within this region.…”

Crystal structure of chloroplast fructose-1,6-bisphosphate aldolase from the green algaChlamydomonas reinhardtii

“…This site is a strongly electropositive pocket, containing one or two sulfate ions captured in the crystallization solution and several water molecules (figure 3). Active site of class I aldolase have already been described structurally and chemically in various organisms (Dalby, Dauter et al 1999) (St-Jean, Lafrance-Vanasse et al 2005) (Lafrance-Vanasse and Sygusch 2007) (Gardberg, Abendroth et al 2011) (Gardberg, Sankaran et al 2011). Eleven catalytic residues described in these articles are present in Chlamydomonas reinhardtii FBA3 at the following positions: Asp52, Ser54, Thr57, Lys125, Lys164, Arg166, Glu204, Glu206, Lys246, Ser288, and Arg318.…”

“…Ser54, Thr57, and Ser64 are localized on the α-helix 2 which is a key element of the catalytic pocket. In fact, Ser54 and Thr57 are part of the amino acids needed to accommodate the substrate in the active site (Dalby, Dauter et al 1999) (St-Jean, Lafrance-Vanasse et al 2005) (Gardberg, Abendroth et al 2011). Phosphorylation of these residues could interfere with the fixation of the substrate both via steric and charge obstructions, supporting a negative regulatory mechanism widely conserved as these residues are conserved from photosynthetic organisms to animals, archaea, and bacteria (supplementary figure 3).…”

Expression of cytosolic and thiolated proteome of Musca domestica larvae under oxidative challenge

“…There was a 6.14 fold increase (Spot J, Figure 1) in expression of fructose bisphosphate aldolase in treated samples. The enzyme catalyzes the forward reaction of aldol splitting of fructose 1,6-bisphosphate to dihydroxyacetone phosphate to glyceraldehyde 3-phosphate and the reverse reaction (Gardberg et al, 2011). Sequence analysis by Brenner-Holzach (1979) however showed no exposed thiol groups of Drosophila melanogaster's fructose bisphosphate aldolase.…”