n - acetylmuramic Search Results


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  • 93
    Millipore n acetylmuramic acid
    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N <t>-acetylmuramic</t> acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.
    N Acetylmuramic Acid, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Carbosynth n acetylmuramic acid
    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N <t>-acetylmuramic</t> acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.
    N Acetylmuramic Acid, supplied by Carbosynth, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Chem Impex International n acetylmuramic acid
    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N <t>-acetylmuramic</t> acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.
    N Acetylmuramic Acid, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Fisher Scientific n acetylmuramic acid
    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N <t>-acetylmuramic</t> acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.
    N Acetylmuramic Acid, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Bachem n acetylmuramic acid murnac
    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N <t>-acetylmuramic</t> acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.
    N Acetylmuramic Acid Murnac, supplied by Bachem, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology sialidase activity substrate 5 bromo 4 chloro 3 indolyl α d n acetylmuramic acid sodium salt
    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N <t>-acetylmuramic</t> acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.
    Sialidase Activity Substrate 5 Bromo 4 Chloro 3 Indolyl α D N Acetylmuramic Acid Sodium Salt, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Agilent technologies udp n acetylmuramyl pentapeptide
    Simplified pathway for bacterial cell wall synthesis. (a) Diagram of key reactions in peptidoglycan synthesis for bacterial cell wall. Abbreviations: MurA, UDP- N -acetylglucosamine enolpyruvyl transferase; MurB, UDP- N -acetylenolpyruvylglucosamine reductase; MurC, UDP- N -acetylmuramoyl-L-alanine synthetase; MurD, UDP- N <t>-acetylmuramyl-l-alanine:d-glutamate</t> ligase; MurE, UDP- N -acetylmuramoyl-l-alanyl-d-glutamate:meso-2,6-diaminopimelate ligase, MurF, UDP- N -acetylmuramoyl-tripeptide-D-alanyl-D-alanine ligase, MurG, UDP- N -acetylglucosamine- N -acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N -acetylglucosamine transferase; IPP, isopentenyl diphosphate, DMAPP, dimethylallyl diphosphate; FPP, isoprenoid farnesyl diphosphate; FPPS, farnesyl diphosphate synthase; UPP, C 55 isoprenoid undecaprenyl diphosphate; UPPS, undecaprenyl diphosphate synthase; UPPP, undecaprenyl diphosphate phosphatase; UP, undecaprenyl monophosphate; MraY, phospho-MurNAc-pentapeptide translocase; GlcNAc, N -acetylglucosamine; MurNAc, N -acetylmuramic acid; PBPs, penicillin-binding proteins. (b) Detection of final soluble cell wall precursor (UDP- N -acetylmuramyl pentapeptide) inside bacterial cytoplasm. HPLC chromatogram of S . aureus NRS107 (RN4220) treated with 10 × MIC of compound 1 or vancomycin for 30 minutes. After centrifugation, the bacterial pellet was boiled for 30 minutes to release contents present in the bacterial cytoplasm. The lysate was analyzed using HPLC/MS, using a phenyl column, to determine the accumulation of the final soluble precursor in cell wall synthesis, UDP- N -acetylmuramyl pentapeptide (designated by the black arrow).
    Udp N Acetylmuramyl Pentapeptide, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore udp murnac pentapeptide
    Analogues of <t>UDP</t> can serve as initiating substrates for RNAP in vitro . ( A ) Template with acnA promoter (partial sequence around transcription start site) and acnAmod template, +1 position is in bold, 12 nt initially transcribed sequence in RNAImod is underlined. Dinucleotide synthesis on acnA template using either RNAP holo σ70 or holo σ70Δ3.2 , and 500 μM UMP, UDP, UTP, UDP-Glc, UDP-GlcNAc and <t>UDP-MurNAc</t> pentapeptide as initiating substrates and (α 32 P)-CTP as the next nucleotide. ( B ) Plot below the gel represents the ratio of incorporation of indicated substrates by holo σ70Δ3.2 versus holo σ70 , the values are an average of the two independent experiments, error bars represent plus and minus one standard deviation. ( C ) Transcripts initiated with UTP, UDP-Glc, UDP-GlcNAc, are elongated to 12 nt transcript on acnAmod template. ( D ) Relative efficiency of incorporation of 500 μM UDP-GlcNAc in comparison to UTP into dinucleotide product on RNAImod template by WT and mutant RNAPs with aminoacid changes in Rifampicin-binding pocket of β subunit indicated below the plot. The values are an average of the two independent experiments, error bars represent plus and minus one standard deviation.
    Udp Murnac Pentapeptide, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    The Medicines Company udp murnac pentapeptide
    Intracellular accumulation of the soluble cell wall precursor <t>UDP-MurNAc-depsipeptide</t> in VanA-type E. faecium . (A) E. faecium BM4147 was grown in HHD broth (0.002% Tween 80) to an OD 600 of 0.5 and supplemented with 80 mg/liter vancomycin (VAN) to induce
    Udp Murnac Pentapeptide, supplied by The Medicines Company, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore pgn
    Intracellular accumulation of the soluble cell wall precursor <t>UDP-MurNAc-depsipeptide</t> in VanA-type E. faecium . (A) E. faecium BM4147 was grown in HHD broth (0.002% Tween 80) to an OD 600 of 0.5 and supplemented with 80 mg/liter vancomycin (VAN) to induce
    Pgn, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 288 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore peptidoglycan
    Persistence of transgenerational immune priming effects on the increase of immune activity of Manduca sexta offspring after offspring immune challenge by <t>PGN</t> in 21-day-old pupae. A) Increase of phenoloxidase (PO) activity and B) increase of antibacterial (AMP) activity (lysozyme activity equivalent, Micrococcus <t>luteus</t> ) were measured in 1-day-old and 3-day-old adults, i.e. three and five days, respectively, after offspring immune treatment in 21-day-old pupae. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Increase of immune activity was measured as increase = (Activity after PGN treatment of the offspring)/(Mean activity of unchallenged offspring); value 1 is labelled by a line that indicates no change in immune activity after offspring challenge. Please note the comparable scales for increases which show the immunity and visualise the strong priming effects on offspring AMP activity, but the lack of effects on PO activity in the offspring. Mean values ± SE are given. N = 9 individuals of each developmental stage from each parental group. Means of absolute data of PGN- and PBS-treated offspring are shown in Table S5 in File S1 . Differences between the parental priming treatments and the time intervals after offspring PGN treatment were compared by two-way ANOVA ( Table 3 ) and post hoc analysis Tukey tests (Table S7 in File S1 ). Statistical evaluation of priming effects on the persistence of immunity after offspring immune challenge by PBS is shown in Table S8 in File S1 .
    Peptidoglycan, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 321 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore muramyldipeptide
    Persistence of transgenerational immune priming effects on the increase of immune activity of Manduca sexta offspring after offspring immune challenge by <t>PGN</t> in 21-day-old pupae. A) Increase of phenoloxidase (PO) activity and B) increase of antibacterial (AMP) activity (lysozyme activity equivalent, Micrococcus <t>luteus</t> ) were measured in 1-day-old and 3-day-old adults, i.e. three and five days, respectively, after offspring immune treatment in 21-day-old pupae. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Increase of immune activity was measured as increase = (Activity after PGN treatment of the offspring)/(Mean activity of unchallenged offspring); value 1 is labelled by a line that indicates no change in immune activity after offspring challenge. Please note the comparable scales for increases which show the immunity and visualise the strong priming effects on offspring AMP activity, but the lack of effects on PO activity in the offspring. Mean values ± SE are given. N = 9 individuals of each developmental stage from each parental group. Means of absolute data of PGN- and PBS-treated offspring are shown in Table S5 in File S1 . Differences between the parental priming treatments and the time intervals after offspring PGN treatment were compared by two-way ANOVA ( Table 3 ) and post hoc analysis Tukey tests (Table S7 in File S1 ). Statistical evaluation of priming effects on the persistence of immunity after offspring immune challenge by PBS is shown in Table S8 in File S1 .
    Muramyldipeptide, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N -acetylmuramic acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.

    Journal: Biochimica et Biophysica Acta

    Article Title: Characterisation of the selective binding of antibiotics vancomycin and teicoplanin by the VanS receptor regulating type A vancomycin resistance in the enterococci

    doi: 10.1016/j.bbagen.2017.05.011

    Figure Lengend Snippet: Activity of VanS A . (A) In the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N -acetylmuramic acid, N -acetylglucosamine). (B) Activity profiles of VanS A in the absence and presence of vancomycin and teicoplanin over 40 min. Standard deviations calculated for each condition ( n = 3) and plotted as error bars.

    Article Snippet: 2.1 Materials Vancomycin, teicoplanin, N-Acetyl-D-glucosamine, N-Acetylmuramic acid, D-Ala-D-Ala, Ala-D-γ-Glu-Lys-D-Ala-D-Ala all purchased from Sigma-Aldrich (UK).

    Techniques: Activity Assay

    Characterisation of ligand binding in the near-UV region (260–230 nm). Difference spectrum of (A) VanS A in the presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N-acetylmuramic acid, D-Ala-D-Ala and Ala-D-y-Glu-Lys-D-Ala-D-Ala); (B) titration of vancomycin and (C) titration of teicoplanin. Binding constant (k d ) calculated from change in CD (mdeg) at (B) 286 nm and (C) 292 nm, plotting against respective concentration (M) of GPA and fitting with a (B) Hill1 or (C) BidoseResponse function. Standard deviation ( n = 4) shown by error bars.

    Journal: Biochimica et Biophysica Acta

    Article Title: Characterisation of the selective binding of antibiotics vancomycin and teicoplanin by the VanS receptor regulating type A vancomycin resistance in the enterococci

    doi: 10.1016/j.bbagen.2017.05.011

    Figure Lengend Snippet: Characterisation of ligand binding in the near-UV region (260–230 nm). Difference spectrum of (A) VanS A in the presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N-acetylmuramic acid, D-Ala-D-Ala and Ala-D-y-Glu-Lys-D-Ala-D-Ala); (B) titration of vancomycin and (C) titration of teicoplanin. Binding constant (k d ) calculated from change in CD (mdeg) at (B) 286 nm and (C) 292 nm, plotting against respective concentration (M) of GPA and fitting with a (B) Hill1 or (C) BidoseResponse function. Standard deviation ( n = 4) shown by error bars.

    Article Snippet: 2.1 Materials Vancomycin, teicoplanin, N-Acetyl-D-glucosamine, N-Acetylmuramic acid, D-Ala-D-Ala, Ala-D-γ-Glu-Lys-D-Ala-D-Ala all purchased from Sigma-Aldrich (UK).

    Techniques: Ligand Binding Assay, Titration, Binding Assay, Concentration Assay, Standard Deviation

    Ligand binding screens of VanS A in the far-UV region (190-260 nm). Difference spectrum of VanS A in the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N-acetylmuramic acid, D-Ala-D-Ala and Ala-D-y-Glu-Lys-D-Ala-D-Ala).

    Journal: Biochimica et Biophysica Acta

    Article Title: Characterisation of the selective binding of antibiotics vancomycin and teicoplanin by the VanS receptor regulating type A vancomycin resistance in the enterococci

    doi: 10.1016/j.bbagen.2017.05.011

    Figure Lengend Snippet: Ligand binding screens of VanS A in the far-UV region (190-260 nm). Difference spectrum of VanS A in the absence and presence of 5-fold molar equivalent of compounds (vancomycin, teicoplanin, N-acetylmuramic acid, D-Ala-D-Ala and Ala-D-y-Glu-Lys-D-Ala-D-Ala).

    Article Snippet: 2.1 Materials Vancomycin, teicoplanin, N-Acetyl-D-glucosamine, N-Acetylmuramic acid, D-Ala-D-Ala, Ala-D-γ-Glu-Lys-D-Ala-D-Ala all purchased from Sigma-Aldrich (UK).

    Techniques: Ligand Binding Assay

    Simplified pathway for bacterial cell wall synthesis. (a) Diagram of key reactions in peptidoglycan synthesis for bacterial cell wall. Abbreviations: MurA, UDP- N -acetylglucosamine enolpyruvyl transferase; MurB, UDP- N -acetylenolpyruvylglucosamine reductase; MurC, UDP- N -acetylmuramoyl-L-alanine synthetase; MurD, UDP- N -acetylmuramyl-l-alanine:d-glutamate ligase; MurE, UDP- N -acetylmuramoyl-l-alanyl-d-glutamate:meso-2,6-diaminopimelate ligase, MurF, UDP- N -acetylmuramoyl-tripeptide-D-alanyl-D-alanine ligase, MurG, UDP- N -acetylglucosamine- N -acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N -acetylglucosamine transferase; IPP, isopentenyl diphosphate, DMAPP, dimethylallyl diphosphate; FPP, isoprenoid farnesyl diphosphate; FPPS, farnesyl diphosphate synthase; UPP, C 55 isoprenoid undecaprenyl diphosphate; UPPS, undecaprenyl diphosphate synthase; UPPP, undecaprenyl diphosphate phosphatase; UP, undecaprenyl monophosphate; MraY, phospho-MurNAc-pentapeptide translocase; GlcNAc, N -acetylglucosamine; MurNAc, N -acetylmuramic acid; PBPs, penicillin-binding proteins. (b) Detection of final soluble cell wall precursor (UDP- N -acetylmuramyl pentapeptide) inside bacterial cytoplasm. HPLC chromatogram of S . aureus NRS107 (RN4220) treated with 10 × MIC of compound 1 or vancomycin for 30 minutes. After centrifugation, the bacterial pellet was boiled for 30 minutes to release contents present in the bacterial cytoplasm. The lysate was analyzed using HPLC/MS, using a phenyl column, to determine the accumulation of the final soluble precursor in cell wall synthesis, UDP- N -acetylmuramyl pentapeptide (designated by the black arrow).

    Journal: PLoS ONE

    Article Title: Bacteriological profiling of diphenylureas as a novel class of antibiotics against methicillin-resistant Staphylococcus aureus

    doi: 10.1371/journal.pone.0182821

    Figure Lengend Snippet: Simplified pathway for bacterial cell wall synthesis. (a) Diagram of key reactions in peptidoglycan synthesis for bacterial cell wall. Abbreviations: MurA, UDP- N -acetylglucosamine enolpyruvyl transferase; MurB, UDP- N -acetylenolpyruvylglucosamine reductase; MurC, UDP- N -acetylmuramoyl-L-alanine synthetase; MurD, UDP- N -acetylmuramyl-l-alanine:d-glutamate ligase; MurE, UDP- N -acetylmuramoyl-l-alanyl-d-glutamate:meso-2,6-diaminopimelate ligase, MurF, UDP- N -acetylmuramoyl-tripeptide-D-alanyl-D-alanine ligase, MurG, UDP- N -acetylglucosamine- N -acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N -acetylglucosamine transferase; IPP, isopentenyl diphosphate, DMAPP, dimethylallyl diphosphate; FPP, isoprenoid farnesyl diphosphate; FPPS, farnesyl diphosphate synthase; UPP, C 55 isoprenoid undecaprenyl diphosphate; UPPS, undecaprenyl diphosphate synthase; UPPP, undecaprenyl diphosphate phosphatase; UP, undecaprenyl monophosphate; MraY, phospho-MurNAc-pentapeptide translocase; GlcNAc, N -acetylglucosamine; MurNAc, N -acetylmuramic acid; PBPs, penicillin-binding proteins. (b) Detection of final soluble cell wall precursor (UDP- N -acetylmuramyl pentapeptide) inside bacterial cytoplasm. HPLC chromatogram of S . aureus NRS107 (RN4220) treated with 10 × MIC of compound 1 or vancomycin for 30 minutes. After centrifugation, the bacterial pellet was boiled for 30 minutes to release contents present in the bacterial cytoplasm. The lysate was analyzed using HPLC/MS, using a phenyl column, to determine the accumulation of the final soluble precursor in cell wall synthesis, UDP- N -acetylmuramyl pentapeptide (designated by the black arrow).

    Article Snippet: UDP-N -acetylmuramyl-pentapeptide was measured using an Agilent High Performance Liquid Chromatography coupled to a time-of-flight Mass Spectrometer (HPLC-MS).

    Techniques: Binding Assay, High Performance Liquid Chromatography, Centrifugation, Mass Spectrometry

    Analogues of UDP can serve as initiating substrates for RNAP in vitro . ( A ) Template with acnA promoter (partial sequence around transcription start site) and acnAmod template, +1 position is in bold, 12 nt initially transcribed sequence in RNAImod is underlined. Dinucleotide synthesis on acnA template using either RNAP holo σ70 or holo σ70Δ3.2 , and 500 μM UMP, UDP, UTP, UDP-Glc, UDP-GlcNAc and UDP-MurNAc pentapeptide as initiating substrates and (α 32 P)-CTP as the next nucleotide. ( B ) Plot below the gel represents the ratio of incorporation of indicated substrates by holo σ70Δ3.2 versus holo σ70 , the values are an average of the two independent experiments, error bars represent plus and minus one standard deviation. ( C ) Transcripts initiated with UTP, UDP-Glc, UDP-GlcNAc, are elongated to 12 nt transcript on acnAmod template. ( D ) Relative efficiency of incorporation of 500 μM UDP-GlcNAc in comparison to UTP into dinucleotide product on RNAImod template by WT and mutant RNAPs with aminoacid changes in Rifampicin-binding pocket of β subunit indicated below the plot. The values are an average of the two independent experiments, error bars represent plus and minus one standard deviation.

    Journal: Nucleic Acids Research

    Article Title: Bacterial RNA polymerase caps RNA with various cofactors and cell wall precursors

    doi: 10.1093/nar/gkx452

    Figure Lengend Snippet: Analogues of UDP can serve as initiating substrates for RNAP in vitro . ( A ) Template with acnA promoter (partial sequence around transcription start site) and acnAmod template, +1 position is in bold, 12 nt initially transcribed sequence in RNAImod is underlined. Dinucleotide synthesis on acnA template using either RNAP holo σ70 or holo σ70Δ3.2 , and 500 μM UMP, UDP, UTP, UDP-Glc, UDP-GlcNAc and UDP-MurNAc pentapeptide as initiating substrates and (α 32 P)-CTP as the next nucleotide. ( B ) Plot below the gel represents the ratio of incorporation of indicated substrates by holo σ70Δ3.2 versus holo σ70 , the values are an average of the two independent experiments, error bars represent plus and minus one standard deviation. ( C ) Transcripts initiated with UTP, UDP-Glc, UDP-GlcNAc, are elongated to 12 nt transcript on acnAmod template. ( D ) Relative efficiency of incorporation of 500 μM UDP-GlcNAc in comparison to UTP into dinucleotide product on RNAImod template by WT and mutant RNAPs with aminoacid changes in Rifampicin-binding pocket of β subunit indicated below the plot. The values are an average of the two independent experiments, error bars represent plus and minus one standard deviation.

    Article Snippet: Materials ATP and UTP were from GE Healthcare; AMP, ADP, NAD+, NADH, NADP+, FAD, UMP, UDP, UDG-Glc and UDP-GlcNAc were from Sigma Aldrich; UDP-MurNAc pentapeptide was a kind gift from Prof. Vollmer.

    Techniques: In Vitro, Sequencing, Gas Chromatography, Standard Deviation, Mutagenesis, Binding Assay

    ADP-related cellular cofactors are utilized by RNAP for initiation of transcription. ( A ) Structures of ATP, NAD + , NADP + , FAD, UDP, UDP-Glc (GlcUDP), UDP-GlcNAc (GlcNAcUDP) and UDP-MurNAc pentapeptide (5AAMurNAcUDP). ( B ) Templates (partial sequence of non-template strand around transcription start site) used for in vitro experiments containing T7A1, RNAI promoters and RNAI template with modified initially transcribed sequence, RNAImod. +1 position for all templates is in bold, −1 position is framed, 9 nt initially transcribed sequence in RNAImod is underlined. ( C ) Initial transcript synthesis on RNAImod template using RNAP holo σ70 , holo σS and holo σ70Δ3.2 and 500μM ATP, NAD + , NADH, NADP + and FAD as initiating substrates and (α 32 P)-CTP as the next nucleotide. ( D ) Plot reflecting incorporation efficiencies for alternative substrates in percentage from efficiency of ATP incorporation for RNAP holo σ70 , holo σS and holo σ70Δ3.2 on RNAImod, the values are an average of the three independent experiments, error bars represent plus and minus one standard deviation.

    Journal: Nucleic Acids Research

    Article Title: Bacterial RNA polymerase caps RNA with various cofactors and cell wall precursors

    doi: 10.1093/nar/gkx452

    Figure Lengend Snippet: ADP-related cellular cofactors are utilized by RNAP for initiation of transcription. ( A ) Structures of ATP, NAD + , NADP + , FAD, UDP, UDP-Glc (GlcUDP), UDP-GlcNAc (GlcNAcUDP) and UDP-MurNAc pentapeptide (5AAMurNAcUDP). ( B ) Templates (partial sequence of non-template strand around transcription start site) used for in vitro experiments containing T7A1, RNAI promoters and RNAI template with modified initially transcribed sequence, RNAImod. +1 position for all templates is in bold, −1 position is framed, 9 nt initially transcribed sequence in RNAImod is underlined. ( C ) Initial transcript synthesis on RNAImod template using RNAP holo σ70 , holo σS and holo σ70Δ3.2 and 500μM ATP, NAD + , NADH, NADP + and FAD as initiating substrates and (α 32 P)-CTP as the next nucleotide. ( D ) Plot reflecting incorporation efficiencies for alternative substrates in percentage from efficiency of ATP incorporation for RNAP holo σ70 , holo σS and holo σ70Δ3.2 on RNAImod, the values are an average of the three independent experiments, error bars represent plus and minus one standard deviation.

    Article Snippet: Materials ATP and UTP were from GE Healthcare; AMP, ADP, NAD+, NADH, NADP+, FAD, UMP, UDP, UDG-Glc and UDP-GlcNAc were from Sigma Aldrich; UDP-MurNAc pentapeptide was a kind gift from Prof. Vollmer.

    Techniques: Gas Chromatography, Sequencing, In Vitro, Modification, Standard Deviation

    Intracellular accumulation of the soluble cell wall precursor UDP-MurNAc-depsipeptide in VanA-type E. faecium . (A) E. faecium BM4147 was grown in HHD broth (0.002% Tween 80) to an OD 600 of 0.5 and supplemented with 80 mg/liter vancomycin (VAN) to induce

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Structural Variations of the Cell Wall Precursor Lipid II and Their Influence on Binding and Activity of the Lipoglycopeptide Antibiotic Oritavancin

    doi: 10.1128/AAC.02663-14

    Figure Lengend Snippet: Intracellular accumulation of the soluble cell wall precursor UDP-MurNAc-depsipeptide in VanA-type E. faecium . (A) E. faecium BM4147 was grown in HHD broth (0.002% Tween 80) to an OD 600 of 0.5 and supplemented with 80 mg/liter vancomycin (VAN) to induce

    Article Snippet: We thank Michaele Josten for performing mass spectrometry and Inge Luhmer-Becker for preparation of the UDP-MurNAc-pentapeptide, Greg Moeck and Adel Rafai Far for discussion and reading of the manuscript, and The Medicines Company for providing oritavancin, des-oritavancin, and chloroeremomycin.

    Techniques:

    Persistence of transgenerational immune priming effects on the increase of immune activity of Manduca sexta offspring after offspring immune challenge by PGN in 21-day-old pupae. A) Increase of phenoloxidase (PO) activity and B) increase of antibacterial (AMP) activity (lysozyme activity equivalent, Micrococcus luteus ) were measured in 1-day-old and 3-day-old adults, i.e. three and five days, respectively, after offspring immune treatment in 21-day-old pupae. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Increase of immune activity was measured as increase = (Activity after PGN treatment of the offspring)/(Mean activity of unchallenged offspring); value 1 is labelled by a line that indicates no change in immune activity after offspring challenge. Please note the comparable scales for increases which show the immunity and visualise the strong priming effects on offspring AMP activity, but the lack of effects on PO activity in the offspring. Mean values ± SE are given. N = 9 individuals of each developmental stage from each parental group. Means of absolute data of PGN- and PBS-treated offspring are shown in Table S5 in File S1 . Differences between the parental priming treatments and the time intervals after offspring PGN treatment were compared by two-way ANOVA ( Table 3 ) and post hoc analysis Tukey tests (Table S7 in File S1 ). Statistical evaluation of priming effects on the persistence of immunity after offspring immune challenge by PBS is shown in Table S8 in File S1 .

    Journal: PLoS ONE

    Article Title: Parental Legacy in Insects: Variation of Transgenerational Immune Priming during Offspring Development

    doi: 10.1371/journal.pone.0063392

    Figure Lengend Snippet: Persistence of transgenerational immune priming effects on the increase of immune activity of Manduca sexta offspring after offspring immune challenge by PGN in 21-day-old pupae. A) Increase of phenoloxidase (PO) activity and B) increase of antibacterial (AMP) activity (lysozyme activity equivalent, Micrococcus luteus ) were measured in 1-day-old and 3-day-old adults, i.e. three and five days, respectively, after offspring immune treatment in 21-day-old pupae. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Increase of immune activity was measured as increase = (Activity after PGN treatment of the offspring)/(Mean activity of unchallenged offspring); value 1 is labelled by a line that indicates no change in immune activity after offspring challenge. Please note the comparable scales for increases which show the immunity and visualise the strong priming effects on offspring AMP activity, but the lack of effects on PO activity in the offspring. Mean values ± SE are given. N = 9 individuals of each developmental stage from each parental group. Means of absolute data of PGN- and PBS-treated offspring are shown in Table S5 in File S1 . Differences between the parental priming treatments and the time intervals after offspring PGN treatment were compared by two-way ANOVA ( Table 3 ) and post hoc analysis Tukey tests (Table S7 in File S1 ). Statistical evaluation of priming effects on the persistence of immunity after offspring immune challenge by PBS is shown in Table S8 in File S1 .

    Article Snippet: Both male and female pupae of the parental generation were subjected to an injection of peptidoglycan extracted from Micrococcus luteus (PGN, Sigma 53243).

    Techniques: Activity Assay, Injection

    Transgenerational immune priming effects on the increase of immune activity of Manduca sexta offspring larvae and pupae one day after offspring immune challenge by PGN. A) Increase of phenoloxidase (PO) activity and B) increase of antibacterial (AMP) activity (lysozyme activity equivalent, Micrococcus luteus ) were measured in 4 th instar larvae and 22-day-old pupae one day after offspring immune treatment. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Increase of immune activity was measured as increase = (Activity after PGN treatment of the offspring)/(Mean activity of unchallenged offspring); value 1 is labelled by a line that indicates no change in immune activity after offspring challenge. Please note the comparable scales for increases which show the immunity and visualise the strong priming effects on offspring AMP activity, but the lack of effects on PO activity in the offspring. Mean values ± SE are given. N = 9 individuals of each developmental stage from each parental group. Means of absolute data of PGN- and PBS-treated offspring are shown in Table S5 in File S1 . Differences between the parental priming treatments and the offspring developmental stages were compared by 2-way-ANOVA ( Table 2 ). Statistical evaluation of priming effects on the increase of immunity after offspring immune challenge by PBS is shown in Table S6 in File S1 .

    Journal: PLoS ONE

    Article Title: Parental Legacy in Insects: Variation of Transgenerational Immune Priming during Offspring Development

    doi: 10.1371/journal.pone.0063392

    Figure Lengend Snippet: Transgenerational immune priming effects on the increase of immune activity of Manduca sexta offspring larvae and pupae one day after offspring immune challenge by PGN. A) Increase of phenoloxidase (PO) activity and B) increase of antibacterial (AMP) activity (lysozyme activity equivalent, Micrococcus luteus ) were measured in 4 th instar larvae and 22-day-old pupae one day after offspring immune treatment. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Increase of immune activity was measured as increase = (Activity after PGN treatment of the offspring)/(Mean activity of unchallenged offspring); value 1 is labelled by a line that indicates no change in immune activity after offspring challenge. Please note the comparable scales for increases which show the immunity and visualise the strong priming effects on offspring AMP activity, but the lack of effects on PO activity in the offspring. Mean values ± SE are given. N = 9 individuals of each developmental stage from each parental group. Means of absolute data of PGN- and PBS-treated offspring are shown in Table S5 in File S1 . Differences between the parental priming treatments and the offspring developmental stages were compared by 2-way-ANOVA ( Table 2 ). Statistical evaluation of priming effects on the increase of immunity after offspring immune challenge by PBS is shown in Table S6 in File S1 .

    Article Snippet: Both male and female pupae of the parental generation were subjected to an injection of peptidoglycan extracted from Micrococcus luteus (PGN, Sigma 53243).

    Techniques: Activity Assay, Injection

    Transgenerational immune priming effects on immune activity of unchallenged Manduca sexta offspring. A ) Phenoloxidase activity and B ) antibacterial activity (lysozyme activity equivalent, Micrococcus luteus ) were measured during offspring development at the first day of each developmental stage and in 22-day-old pupae. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Means ± SE are given. N = 9 samples of each developmental stage from each parental group. Differences between the parental priming treatments and the offspring developmental stages were compared by 2-way-ANOVA ( Table 1 ) and post-hoc analysis Tukey tests (Table S3 for PO activity, table S4 for antibacterial activity in File S1 ).

    Journal: PLoS ONE

    Article Title: Parental Legacy in Insects: Variation of Transgenerational Immune Priming during Offspring Development

    doi: 10.1371/journal.pone.0063392

    Figure Lengend Snippet: Transgenerational immune priming effects on immune activity of unchallenged Manduca sexta offspring. A ) Phenoloxidase activity and B ) antibacterial activity (lysozyme activity equivalent, Micrococcus luteus ) were measured during offspring development at the first day of each developmental stage and in 22-day-old pupae. Female and male parents received a priming treatment in their pupal stage: Naive) untreated, PBS) control-injected with phosphate buffered saline, PGN) injected with peptidoglycan. If the symbol for offspring of naive parents is not visible, it is overlaid by another symbol. Means ± SE are given. N = 9 samples of each developmental stage from each parental group. Differences between the parental priming treatments and the offspring developmental stages were compared by 2-way-ANOVA ( Table 1 ) and post-hoc analysis Tukey tests (Table S3 for PO activity, table S4 for antibacterial activity in File S1 ).

    Article Snippet: Both male and female pupae of the parental generation were subjected to an injection of peptidoglycan extracted from Micrococcus luteus (PGN, Sigma 53243).

    Techniques: Activity Assay, Injection

    Effects of siRNA on the levels of (A) creatine kinase (CK), (B) lactate dehydrogenase (LDH), (C) interleukin (IL)-6 and (D) tumor necrosis factor (TNF)-α in the culture supernatant of lipopolysaccharide (LPS)/peptidoglycan G (PepG)-treated H9C2 cells. Control, LPS/PepG, LPS/PepG + negative control siRNA (ncRNA), LPS/PepG + siRNA represent the control, LPS/PepG, LPS/PepG + ncRNA and LPS/PepG + siRNA group, respectively. The data are expressed as the means ± SD; n=3 for each group. *P

    Journal: International Journal of Molecular Medicine

    Article Title: Silencing of uncoupling protein 2 by small interfering RNA aggravates mitochondrial dysfunction in cardiomyocytes under septic conditions

    doi: 10.3892/ijmm.2015.2177

    Figure Lengend Snippet: Effects of siRNA on the levels of (A) creatine kinase (CK), (B) lactate dehydrogenase (LDH), (C) interleukin (IL)-6 and (D) tumor necrosis factor (TNF)-α in the culture supernatant of lipopolysaccharide (LPS)/peptidoglycan G (PepG)-treated H9C2 cells. Control, LPS/PepG, LPS/PepG + negative control siRNA (ncRNA), LPS/PepG + siRNA represent the control, LPS/PepG, LPS/PepG + ncRNA and LPS/PepG + siRNA group, respectively. The data are expressed as the means ± SD; n=3 for each group. *P

    Article Snippet: In order to simulate sepsis, some cells were cultured in the presence of 2 μ g/ml lipopolysaccharide (LPS, from Escherichia coli O111:B4; Sigma-Aldrich, St. Louis, MO, USA) plus 20 μ g/ml peptidoglycan G (PepG, from Micrococcus luteus; Sigma-Aldrich).

    Techniques: Negative Control

    Effect of siRNA on mitochondrial membrane potential (MMP or ΔΨm) in H9C2 cells under septic conditions. (A) Scanning of MMP in H9C2 cells using a confocal microscope. Green fluorescence represents the monomeric form of the JC-1 molecule, which appears in the cytosol after mitochondrial membrane depolarization. Red fluorescence reveals the mitochondrial aggregate form of JC-1, indicating MMP (magnification, x40). (B) Evaluation of MMP in H9C2 cells by flow cytometry (FCM). FITC-H, green; PI-H, red. (A and B) Panel a, control group; panel b, lipopolysaccharide (LPS)/peptidoglycan G (PepG) group; panel c, LPS/PepG + negative control siRNA (ncRNA) group; panel d, LPS/PepG + siRNA group; panel e, carbonyl cyanide m-chlorophenylhydrazone (CCCP)-treated cells. (C) The ratio of the red over the green fluorescence intensity by FCM represents the quantitative MMP in each group. Values are expressed as the means ± SD.  * P

    Journal: International Journal of Molecular Medicine

    Article Title: Silencing of uncoupling protein 2 by small interfering RNA aggravates mitochondrial dysfunction in cardiomyocytes under septic conditions

    doi: 10.3892/ijmm.2015.2177

    Figure Lengend Snippet: Effect of siRNA on mitochondrial membrane potential (MMP or ΔΨm) in H9C2 cells under septic conditions. (A) Scanning of MMP in H9C2 cells using a confocal microscope. Green fluorescence represents the monomeric form of the JC-1 molecule, which appears in the cytosol after mitochondrial membrane depolarization. Red fluorescence reveals the mitochondrial aggregate form of JC-1, indicating MMP (magnification, x40). (B) Evaluation of MMP in H9C2 cells by flow cytometry (FCM). FITC-H, green; PI-H, red. (A and B) Panel a, control group; panel b, lipopolysaccharide (LPS)/peptidoglycan G (PepG) group; panel c, LPS/PepG + negative control siRNA (ncRNA) group; panel d, LPS/PepG + siRNA group; panel e, carbonyl cyanide m-chlorophenylhydrazone (CCCP)-treated cells. (C) The ratio of the red over the green fluorescence intensity by FCM represents the quantitative MMP in each group. Values are expressed as the means ± SD. * P

    Article Snippet: In order to simulate sepsis, some cells were cultured in the presence of 2 μ g/ml lipopolysaccharide (LPS, from Escherichia coli O111:B4; Sigma-Aldrich, St. Louis, MO, USA) plus 20 μ g/ml peptidoglycan G (PepG, from Micrococcus luteus; Sigma-Aldrich).

    Techniques: Microscopy, Fluorescence, Flow Cytometry, Cytometry, Negative Control

    Effect of siRNA on intracellular reactive oxygen species (ROS) production, adenosine triphosphate (ATP) levels and mitochondrial DNA (mtDNA) copy numbers. The fluorescence intensity of DCF in H9C2 cells represented the production of intracellular ROS. (A) Changes in ROS production measured as the DCF fluorescence under a fluorescence microscope (magnification, x10); Panels a–e, ROS-positive control cells (Rosup), control, lipopolysaccharide (LPS)/peptidoglycan G (PepG), LPS/PepG + negative control siRNA (ncRNA) and LPS/PepG + siRNA group, respectively. (B) DCF fluorescence was read by a multifunctional microplate reader at an excitation wavelength of 488 nm and at an emission wavelength of 525 nm. (C) Cellular ATP expressed as nmol/mg protein was measured using a firefly luciferase-based ATP assay kit. RLU values were measured by a microplate reader. (D) Relative mtDNA copy numbers were detected by real-time PCR with normalization to 18s RNA. Values are expressed as the means ± SD.  * P

    Journal: International Journal of Molecular Medicine

    Article Title: Silencing of uncoupling protein 2 by small interfering RNA aggravates mitochondrial dysfunction in cardiomyocytes under septic conditions

    doi: 10.3892/ijmm.2015.2177

    Figure Lengend Snippet: Effect of siRNA on intracellular reactive oxygen species (ROS) production, adenosine triphosphate (ATP) levels and mitochondrial DNA (mtDNA) copy numbers. The fluorescence intensity of DCF in H9C2 cells represented the production of intracellular ROS. (A) Changes in ROS production measured as the DCF fluorescence under a fluorescence microscope (magnification, x10); Panels a–e, ROS-positive control cells (Rosup), control, lipopolysaccharide (LPS)/peptidoglycan G (PepG), LPS/PepG + negative control siRNA (ncRNA) and LPS/PepG + siRNA group, respectively. (B) DCF fluorescence was read by a multifunctional microplate reader at an excitation wavelength of 488 nm and at an emission wavelength of 525 nm. (C) Cellular ATP expressed as nmol/mg protein was measured using a firefly luciferase-based ATP assay kit. RLU values were measured by a microplate reader. (D) Relative mtDNA copy numbers were detected by real-time PCR with normalization to 18s RNA. Values are expressed as the means ± SD. * P

    Article Snippet: In order to simulate sepsis, some cells were cultured in the presence of 2 μ g/ml lipopolysaccharide (LPS, from Escherichia coli O111:B4; Sigma-Aldrich, St. Louis, MO, USA) plus 20 μ g/ml peptidoglycan G (PepG, from Micrococcus luteus; Sigma-Aldrich).

    Techniques: Fluorescence, Microscopy, Positive Control, Negative Control, Luciferase, ATP Assay, Real-time Polymerase Chain Reaction