Streptomycin affinity depends on 13 amino acids forming a loop in homology modelled ribosomal S12 protein (rpsL gene) ofLysinibacillus sphaericusDSLS5 associated with marine sponge (Tedania anhelans)

Abstract

Streptomycin, an antibiotic used against microbial infections, inhibits the protein synthesis by binding to ribosomal protein S12, encoded by rpsL12 gene, and associated mutations cause streptomycin resistance. A streptomycin resistant, Lysinibacillus sphaericus DSLS5 (MIC >300 µg/mL for streptomycin), was isolated from a marine sponge (Tedania anhelans). The characterisation of rpsL12 gene showed a region having similarity to long terminal repeat sequences of murine lukemia virus which added 13 amino acids for loop formation in RpsL12; in addition, a K56R mutation which corresponds to K43R mutation present in streptomycin-resistant Escherichia coli is also present. The RpsL12 protein was modelled and compared with that of Lysinibacillus boronitolerans, Escherichia coli and Mycobacterium tuberculosis. The modelled proteins docked with streptomycin indicate compound had less affinity. The effect of loop on streptomycin resistance was analysed by constructing three different models of RpsL12 by, (i) removing both loop and mutation, (ii) removing the loop alone while retaining the mutation and (iii) without mutation having loop. The results showed that the presence of loop causes streptomycin resistance (decreases the affinity), and it further enhanced in the presence of mutation at 56th codon. Further study will help in understanding the evolution of streptomycin resistance in organisms.

Development of a Fluorescence Polarization Assay for Multi-Determination of 10 Aminoglycosides in Pork Muscle Sample Based on Ribosomal Protein S12 and Studying Its Recognition Mechanism

“…VAL38 from β2 (interacting with 7 AGs) and SER40 from β3 (interacting with 6 AGs) constructed the pocket bottom; GYL27 from β1 (interacting with 5 AGs) and TYR28 from β1 (interacting with 6 AGs) constructed one side of the pocket, and GLN42 from β3 (interacting with 5 AGs), LYS43 from β3 (interacting with 5 AGs) and ARG44 from β3 (interacting with 6 AGs) and THR71 from β4 (interacting with 5 AGs) constructed the other side of the pocket. The positions of these contact amino acids were similar to the previous report [ 35 ]. As shown in Figure 1 , the specific binding sites in the 10 AGs were different, and this may be because of their different molecular structures.…”

“…As far as we know, there has been only one study reporting the intermolecular interaction of L. sphaericus RpsL 12 with AGs (streptomycin) [ 35 ]. The results showed that the amino acids at the 40–45 positions constructed the binding pocket, and hydrogen bonds (Thr40, Pro41, Arg42, Lys43, Asn45) and hydrophobic interaction (Pro41 and Pro44) were the main intermolecular forces.…”

“…AGs can bind 16S rRNA helices and ribosomal protein S12 ( RpsL 12) to interfere with the function of the bacterial ribosome and show an antibacterial effect [ 29 , 30 , 31 ]. Furthermore, the amino acid mutations in RpsL 12 have been proven to be the major mechanism of AG-resistant bacterial strains [ 32 , 33 , 34 , 35 ]. Therefore, RpsL 12 is the AG receptor that should recognize and bind to all AG species.…”

A streptomycin resistance marker inH. parasuisbased on site-directed mutations inrpsLgene to perform unmarked in-frame mutations and to verify natural transformation

“…It has been well established in procaryotic organisms that streptomycin hinders the functioning of ribosome by binding to 16S rRNA helices and ribosomal protein S12, encoded by rrs and rpsL , respectively. SM also interacts with decoding site and shifts 16S rRNA helix in the direction of ribosomal protein S12 which in turn induces miscoding by stabilising the closed confirmation of 30S ribosomal subunit ( Suriyanarayanan et al, 2016 ). We directly sequenced the rpsL and rrs genes for wild-type strains Hps32, Hps32 and artificially induced SM-resistant strains.…”

Production of a Natural Dihydropteroate Synthase and Development of a Signal-Amplified Pseudo-Immunoassay for the Determination of Sulfonamides in Pork

“…However, all of the previously used DHPS proteins for the analysis of SAs are the recombinant products that are expressed according to the genes of previous studies, so they should have some differences from the natural DHPS proteins. It is well-known that the production of a natural receptor using a conventional procedure is tedious and expensive, and the obtained product has many uncertainties. , In comparison, the use of photoaffinity-labeled activity-based protein-profiling probe (PAL-ABPP) to produce a natural receptor is simple and cheap, so this type of probe has been used to study the receptors of some small-molecule substances. , In our recent report, a type of PAL-ABPP was synthesized; it was used to capture the natural TetR protein from a tetracycline-resistant bacteria, and the obtained receptor simultaneously recognized all of the tested tetracycline drugs with comparable sensitivities …”