11 research outputs found

    Main muropeptides from <i>L. casei</i> BL23 PG hydrolyzed by Lc-p75 and main products of digestion.

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    a<p>Peak numbers refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032301#pone-0032301-g002" target="_blank">Figure 2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032301#pone.0032301.s004" target="_blank">Figure S3</a>. New peaks obtained after Lc-p75 digestion, are indicated by letters.</p>b<p>Di, disaccharide dipeptide (L-Ala-D-iGln); Tri, disaccharide tripeptide (L-Ala-D-iGln-L-Lys); Tetra, disaccharide tetrapeptide (L-Ala-D-iGln-L-Lys-D-Ala); Disaccharide, GlcNAc-MurNAc; Ac, acetylation on MurNAc, iGln, isoglutamine; N, D-Asn; A, D-Ala; K, L-Lys.</p>c<p>Sodiated molecular ions were the most abundant ones on MALDI-TOF mass spectra for all muropeptides.</p>d<p>Percentage of each peak was calculated as the ratio of the peak area over the sum of areas of all the peaks identified in the corresponding chromatogram (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032301#pone.0032301.s008" target="_blank">Table S3</a>).</p>e<p>ND, non detected.</p

    RP-HPLC separation profile of muropeptides obtained from <i>L. casei</i> BL23 PG.

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    <p>PG was digested by mutanolysin (A) or by mutanolysin and recombinant Lc-p75 (B). Numbers correspond to the main muropeptides which amounts changed between Panel A and B. Letters indicate new peaks present in Panel B and absent in Panel A. Complete annotation of the chromatograms is presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032301#pone.0032301.s004" target="_blank">Figure S3</a>.</p

    Lc-p75 activity on purified muropeptides selected as substrates.

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    a<p>Di, disaccharide dipeptide; Tri, disaccharide tripeptide (L-Ala-D-iGln-L-Lys); Tetra, disaccharide tetrapeptide (L-Ala-D-iGln-L-Lys-D-Ala); Disaccharide, GlcNAc-MurNAc; iGln, isoglutamine; N, Asn; Ac, acetylation on MurNAc.</p>b<p>Similar amounts of each muropeptide were used for each test.</p>c<p>Percentage of each peak was calculated as the ratio of the peak area over the sum of areas of all the peaks identified in the corresponding chromatogram.</p>d<p>Other forms of muropeptides resulting from partial digestion of the substrate.</p

    Detection of Lc-p75 in culture supernatant.

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    <p>Analysis by SDS-PAGE (A, B) and zymogram assay (C, D) of the culture supernatant of wild type BL23 (A, C) and negative mutant (B, D) grown on AOAC medium. Lc-p75 is indicated by an arrow.</p

    Indirect immunofluorescence localization of Strep-tagged Lc-p75 in <i>L. casei</i>.

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    <p>Strep-tagged Lc-p75 was localized in overexpressing strain (A) and in complemented negative mutant (B) with monoclonal antibody directed against Strep-tag as first antibody.</p

    Phenotype comparison between wild type <i>L. casei</i> BL23, Lc-p75-negative mutant and complemented Lc-p75 mutant.

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    <p>Pictures of wild type <i>L. casei</i> (A, C, F, H), Lc-p75-negative mutant (B, D, I) and complemented Lc-p75 mutant (E). Colony morphology (A, B), phase contrast microscopy (C, D, E) fluorescence microscopy with merged FM-4-64 (red) and DAPI (blue) staining (F, G) and transmission electron microscopy (H, I).</p

    Indirect immunofluorescence microscopy (A,B,C&D).

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    <p>Anti-Msp1 (A&B) and anti-Msp2 (C&D) rabbit antisera were used on wild-type (A&C) en <i>msp1</i> mutant cells (B&D). Anti-rabbit IgG antibodies conjugated with Alexa Fluor 488 (Invitrogen) were used to visualize the Msp1 and Msp2 localization on the cells.</p

    Main muropeptides from LGG PG digested with or without Msp1.

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    a<p>Peak numbers refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031588#pone-0031588-g005" target="_blank">Figure 5</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031588#pone.0031588.s001" target="_blank">Figure S1</a>.</p>b<p>Di, disaccharide dipeptide (L-Ala-D-iGln); Tri, disaccharide tripeptide (L-Ala-D-iGln-L-Lys); Tetra, disaccharide tetrapeptide (L-Ala-D-iGln-L-Lys-D-Ala); Disaccharide, GlcNAc-MurNAc; Ac, acetylation, iGln, isoglutamine; N, D-Asn; A, D-Ala; K, L-Lys.</p>c<p>Sodiated molecular ions were the most abundant ones on MALDI-TOF mass spectra for all muropeptides.</p>d<p>Percentage of each peak was calculated as the ratio of the peak area over the sum of areas of all the peaks identified in the corresponding chromatogram (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031588#pone-0031588-g005" target="_blank">Figure 5</a>).</p>e<p>ND, non detected.</p

    RP-HPLC separation profile of muropeptides from LGG digested with mutanolysin (A) and digested with mutanolysin and Msp1 (B).

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    <p>Peak numbers refer to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031588#pone-0031588-t001" target="_blank">Table 1</a>. DS-di, disaccharide dipeptide (GlcNAc-MurNAc-L-Ala-D-Gln). The schematic structure of LGG peptidoglycan and site of cleavage of Msp1 is also represented. GlcNAc, N-acetylglucosamine; MurNAc, N-acetylmuramic acid.</p
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