ABSTRACT
To investigate the function of the pa4079 gene from the opportunistic pathogen Pseudomonas aeruginosa PAO1, we determined its crystal structure and confirmed it to be a NAD(P)-dependent short-chain dehydrogenase/reductase. Structural similarity and activity for a broad range of substrates indicate that PA4079 functions as a carbonyl reductase. Comparison of apo- and holo-PA4079 shows that NADP stabilizes the active site specificity loop, and small molecule binding induces rotation of the Tyr183 side chain by approximately 90° out of the active site. Quantitative real-time PCR results show that pa4079 maintains high expression levels during antibiotic exposure. This work provides a starting point for understanding substrate recognition and selectivity by PA4079, as well as its possible reduction of antimicrobial drugs. DATABASE: Structural data are available in the Protein Data Bank (PDB) under the following accession numbers: apo PA4079 (condition I), 5WQM; apo PA4079 (condition II), 5WQN; PA4079 + NADP (condition I), 5WQO; PA4079 + NADP (condition II), 5WQP.
Subject(s)
Aldehyde Reductase/metabolism , Bacterial Proteins/metabolism , Butyryl-CoA Dehydrogenase/metabolism , Models, Molecular , NADP/metabolism , Pseudomonas aeruginosa/metabolism , Aldehyde Reductase/chemistry , Aldehyde Reductase/genetics , Aldo-Keto Reductases , Amino Acid Sequence , Amino Acid Substitution , Anti-Bacterial Agents/pharmacology , Apoenzymes/chemistry , Apoenzymes/genetics , Apoenzymes/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Binding Sites , Butyryl-CoA Dehydrogenase/chemistry , Butyryl-CoA Dehydrogenase/genetics , Catalytic Domain , Conserved Sequence , Crystallography, X-Ray , Enzyme Stability , Gene Expression Regulation, Bacterial/drug effects , Holoenzymes/chemistry , Holoenzymes/genetics , Holoenzymes/metabolism , Ligands , Mutation , NADP/chemistry , Protein Conformation , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/growth & development , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Sequence Alignment , Structural Homology, Protein , Substrate SpecificityABSTRACT
OBJECTIVE: To study the effect of Suxiao Jiuxinwan on angiogenesis in experimental myocardial infarction rats. METHOD: The animal model was established by ligation of anterior decending coronary artery and the infarction areas as well as microvascular density (MVD) in the marginal infarction area of the myocardial infarction rats were observed. RESULT: Infarction areas in high dose and low dose Suxiao Jiuxinwan groups were significantly different from that of model group (P<0.01), and this effect was similar with the positive model group and had the dosage-dependent speciality. The microvascular density (MVD) of marginal infarction areas in Suxiao Jiuxinwan high and low groups was much more elevated than that in the sham group (P<0.05), and the effect of Suxiao Jiuxinwan high group was similar with that of the positive control group. But the difference between the Suxiao Jiuxinwan high and the low goups was not significant. CONCLUSION: Suxiao Jiuxinwan has the obvious effcet on angiogenesis in eperimental myocardial infarction rats.
