s pneumoniae d39  (ATCC)

Journal: Molecules

Article Title: Shionone-Targeted Pneumolysin to Ameliorate Acute Lung Injury Induced by Streptococcus pneumoniae In Vivo and In Vitro

doi: 10.3390/molecules27196258

Figure Lengend Snippet: Shionone inhibits PLY hemolytic activity and does not affect S. pneumoniae growth at less than 8 μg/mL. ( A ) Shionone chemical structure formula. ( B ) Shionone exhibits a concentration-dependent inhibitory effect on the hemolytic activity of PLY. ( C ) The growth curves of S. pneumoniae were determined under shionone treatment with different concentrations (0 μg/mL, 4 μg/mL and 8 μg/mL). ( D ) The fluorescence images of S. pneumoniae treated with different concentrations of shionone (0 μg/mL, 4 μg/mL, and 8 μg/mL) for 6 h. Green in the image represents live cells, and red represents dead cells. The significance level was expressed as follows: ** p < 0.01.

Article Snippet: The S. pneumoniae strain D39 (ATCC49619) used in this study was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). and cultured according to previous research [ ].

Techniques: Activity Assay, Concentration Assay, Fluorescence

Journal: Molecules

Article Title: Shionone-Targeted Pneumolysin to Ameliorate Acute Lung Injury Induced by Streptococcus pneumoniae In Vivo and In Vitro

doi: 10.3390/molecules27196258

Figure Lengend Snippet: Shionone inhibits virulence of S. pneumoniae by targeting the oligomerization of PLY. ( A ) The oligomerization of PLY was detected by western blotting treated with or without shionone. ( B ) Densitometric analysis of PLY oligomerization with different concentrations of shionone treatment. ( C ) The expression level of PLY determined after S. pneumoniae was cocultured with different concentrations of shionone for 6 h. ( D , E ) Shionone interacts with the active pocket of PLY and targets amino acid sites ASP-59, ILE-60, THR-57, PHE-344, and ASN-346. The significance level was expressed as follows: ** p < 0.01.

Article Snippet: The S. pneumoniae strain D39 (ATCC49619) used in this study was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). and cultured according to previous research [ ].

Techniques: Western Blot, Expressing

Journal: Molecules

Article Title: Shionone-Targeted Pneumolysin to Ameliorate Acute Lung Injury Induced by Streptococcus pneumoniae In Vivo and In Vitro

doi: 10.3390/molecules27196258

Figure Lengend Snippet: Shionone alleviates mice pneumonia injury induced by S. pneumoniae . ( A , B ) Gross lung tissue lesions and hematoxylin/eosin staining analysis after infected for 48 h with or without 50 mg/kg shionone treatment. ( C ) Lung colony count of the control, model, and treatment groups after infection for 48 h. The significance level was expressed as follows: ** p < 0.01; NS, no significant difference.

Article Snippet: The S. pneumoniae strain D39 (ATCC49619) used in this study was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). and cultured according to previous research [ ].

Techniques: Staining, Infection, Control

Journal: Science Advances

Article Title: The structural basis of bacterial manganese import

doi: 10.1126/sciadv.abg3980

Figure Lengend Snippet: ( A ) F100S PsaB 2 C 2 -Fab 2 complex (space group C 222 1 ; PsaC, blue and lavender; PsaB, hot and light pink; Fabs, gray). ( B ) F100S PsaB 2 C 2 structure [space group P 1; colored as in (A)]. ( C ) Superposition of F100S PsaB 2 C 2 -Fab 2 and F100S PsaB 2 C 2 structures [ F100S PsaB 2 C 2 -Fab 2 colored as in (B); F100S PsaB 2 C 2 in gray]. Fabs omitted for clarity. ( D ) MD snapshot of WT PsaB 2 C 2 [colored as in (A)] in a PG and CDL membrane. Headgroups of lipid species (PG, yellow; CDL species, orange) are shown as spheres with tails hidden. ( E ) ATPase activity of reconstituted WT PsaBC and F100S PsaBC with Mn 2+ and/or PsaA. Data represent means (± SEM) from at least four independent experiments. ( F ) Growth of WT S. pneumoniae (D39 rpsl + ), the isogenic mutant, and complemented strains (Ω) in Mn 2+ -limited media. Data represent means (± SD) absorbance measurements from three independent experiments. ( G ) Mn 2+ accumulation of S. pneumoniae strains in Mn 2+ -limited media. Data represent means (± SD) from three independent experiments. Statistical significance was determined by one-way analysis of variance (ANOVA) with Tukey posttest P > 0.05, not significant (ns); **** P < 0.0001. ( H ) PsaC (from F100S PsaB 2 C 2 ) with helices represented as cylinders. One monomer is colored gray, the other is colored from blue (N terminus) to red (C terminus). ( I ) Topology of PsaC monomers with helices numbered (AH, amphipathic helix; EG, extracellular gate; CH, coupling helix). Colors correspond to (H).

Article Snippet: The S. pneumoniae D39 genome (National Center for Biotechnology Information identifier NC_008533) served as the reference genome to determine the presence, amino acid sequence, and alignment of spd_1461 ( psaB ), spd_1462 ( psaC ), and spd_1463 ( psaA ) across the 20,020 clinical isolates using the screen_assembly script ( ) and BLASTN v2.9.0 with parameters of 80% coverage and 80% identity.

Techniques: Membrane, Activity Assay, Mutagenesis

Journal: Science Advances

Article Title: The structural basis of bacterial manganese import

doi: 10.1126/sciadv.abg3980

Figure Lengend Snippet: ( A ) Structural elements composing the translocation pathway with Phe 121 , Leu 124 , Phe 125 , Leu 104 , His 50 , and Asp 46 shown as sticks. Residue conservation (65 TMDs from 52 characterized transporters; fig. S6) mapped to PsaC (<50%, white; 50 to 100%, white to blue). ( B ) Volume of the F100S PsaB 2 C 2 translocation pathway visualized in purple; arrow denotes 0.7-Å closure. ( C ) π-π stacking between Phe 121 and Phe 125 . ( D ) MD analysis of WT PsaB 2 C 2 in a solvated membrane (8 × 500-ns simulations). Representative starting (left) and final (right) frames with time-dependent water occupancy displayed as a heat matrix (middle) aligned to the structure (dashed lines). PsaB (gray cartoon), PsaC (TM2 and TM4; black ribbons), and water molecules (blue sticks). ( E ) Representation of PsaB 2 C 2 with negatively charged surfaces in red, positive in blue, and uncharged in white. Cutaway reveals the CS (indicated by arrow). ( F ) ATPase activity of reconstituted D46A PsaBCA and H50A PsaBCA in the presence or absence of Mn 2+ . Data represent means (± SEM) from three independent experiments. ( G ) Growth of WT S. pneumoniae (D39 rpsl + ) and the mutant strains in Mn 2+ -limited medium. Data represent means (± SD) absorbance measurements from three independent experiments. ( H ) Mn 2+ accumulation of S. pneumoniae strains in Mn 2+ -limited media. Data represent means (± SD) from three independent experiments. Statistical significance was determined by one-way ANOVA with Tukey posttest [ P > 0.05, not significant (ns); **** P < 0.0001].

Article Snippet: The S. pneumoniae D39 genome (National Center for Biotechnology Information identifier NC_008533) served as the reference genome to determine the presence, amino acid sequence, and alignment of spd_1461 ( psaB ), spd_1462 ( psaC ), and spd_1463 ( psaA ) across the 20,020 clinical isolates using the screen_assembly script ( ) and BLASTN v2.9.0 with parameters of 80% coverage and 80% identity.

Techniques: Translocation Assay, Residue, Membrane, Activity Assay, Mutagenesis

Journal: Journal of the American Society for Mass Spectrometry

Article Title: Top-Down LESA Mass Spectrometry Protein Analysis of Gram-Positive and Gram-Negative Bacteria

doi: 10.1007/s13361-017-1718-8

Figure Lengend Snippet: Liquid extraction surface analysis (LESA) mass spectra of seven bacterial strains: Escherichia coli K-12, Escherichia coli BL21 mCherry, Pseudomonas aeruginosa PS1054, Staphylococcus aureus MSSA476, Streptococcus pneumoniae D39, Streptococcus oralis ATCC 35037, and Streptococcus gordonii ATCC 35105. All mass spectra were acquired immediately following incubation of the colonies at 37 °C, with use of the 40:60:1 acetonitrile–water–formic acid solvent system for Gram-negative bacteria and the 50:45:5 variant for Gram-positive bacteria. The streptococci were incubated under semianaerobic conditions for optimum growth; the remaining strains were grown in open air

Article Snippet: As an additional challenge, colonies of three species of Streptococcus — S. pneumoniae D39, S. oralis ATCC 35037, and S. gordonii ATCC 35105—were subjected to LESA MS characterization.

Techniques: Extraction, Incubation, Solvent, Bacteria, Variant Assay

Journal: Journal of the American Society for Mass Spectrometry

Article Title: Top-Down LESA Mass Spectrometry Protein Analysis of Gram-Positive and Gram-Negative Bacteria

doi: 10.1007/s13361-017-1718-8

Figure Lengend Snippet: Proteins Identified by Liquid Extraction Surface Analysis Mass Spectrometry in Sampled Colonies

Article Snippet: As an additional challenge, colonies of three species of Streptococcus — S. pneumoniae D39, S. oralis ATCC 35037, and S. gordonii ATCC 35105—were subjected to LESA MS characterization.

Techniques: Extraction, Mass Spectrometry, Molecular Weight, Incubation