6fux

Structure of aminoglycoside phosphotransferase APH(3)-Id from Streptomyces rimosus ATCC10970 in complex with ADP and streptomycin

spinqualitylabelsall models 6fux, resolution 1.65Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 6fux is a 1 chain structure with sequence from Strr1. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: ADP, GOL, SRY Related: 6fuc Gene: SRIM_08058 (STRR1) Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Publication Abstract from PubMed In this study, we identified a new gene (aph(3)-Id) coding for a streptomycin phosphotransferase by using phylogenetic comparative analysis of the genome of the oxytetracycline-producing strain Streptomyces rimosus ATCC 10970. Cloning the aph(3)-Id gene in E.coli and inducing its expression led to an increase in the minimum inhibitory concentration of the recombinant E.coli strain to streptomycin reaching 350mug/ml. To evaluate the phosphotransferase activity of the recombinant protein APH(3)-Id we carried out thin-layer chromatography of the putative (32)P-labeled streptomycin phosphate. We also performed a spectrophotometric analysis to determine the production of ADP coupled to NADH oxidation. Here are the kinetic parameters of the streptomycin phosphotransferase APH(3)-Id: Km 80.4muM, Vmax 6.45mumol/min/mg and kcat 1.73 s(-1). We demonstrated for the first time the ability of the aminoglycoside phototransferase (APH(3)-Id) to undergo autophosphorylation in vitro. The 3D structures of APH(3)-Id in its unliganded state and in ternary complex with streptomycin and ADP were obtained. The structure of the ternary complex is the first example of this class of enzymes with bound streptomycin. Comparison of the obtained structures with those of other aminoglycoside phosphotransferases revealed peculiar structure of the substrate-binding pocket reflecting its specificity to a particular antibiotic. Identification, functional and structural characterization of novel aminoglycoside phosphotransferase APH(3)-Id from Streptomyces rimosus subsp. rimosus ATCC 10970.,Alekseeva MG, Boyko KM, Nikolaeva AY, Mavletova DA, Rudakova NN, Zakharevich NV, Korzhenevskiy DA, Ziganshin RH, Popov VO, Danilenko VN Arch Biochem Biophys. 2019 Jun 26;671:111-122. doi: 10.1016/j.abb.2019.06.008. PMID:31251922 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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