Journal:

Article Title: The Antifungal Protein from Aspergillus giganteus Causes Membrane Permeabilization

doi: 10.1128/AAC.47.2.588-593.2003

Figure Lengend Snippet: Effects of different ions on the MIC of AFP for A. niger. The MIC of AFP for A. niger was tested in the presence of different amounts of KH2PO4 (○), KCl (•), and NaCl (▴). The results of a representative experiment are shown.

Article Snippet: Aspergillus niger ATCC 9029 , 1.

Techniques:

Journal:

Article Title: The Antifungal Protein from Aspergillus giganteus Causes Membrane Permeabilization

doi: 10.1128/AAC.47.2.588-593.2003

Figure Lengend Snippet: Detection of AFP-induced SYTOX Green uptake by fluorescence microscopy. A. niger (A and B) and P. chrysogenum (C and D) were treated with 10 and 100 μg of AFP, respectively, in the presence of 0.2 μM SYTOX Green. After 1 h of incubation, SYTOX Green uptake was detected by fluorescence microscopy. (A and C) Fluorescence microscopy; (B and D) light-field microscopy. Bars, 15 μm.

Article Snippet: Aspergillus niger ATCC 9029 , 1.

Techniques: Fluorescence, Microscopy, Incubation

Journal:

Article Title: The Antifungal Protein from Aspergillus giganteus Causes Membrane Permeabilization

doi: 10.1128/AAC.47.2.588-593.2003

Figure Lengend Snippet: Detection of AFP-induced SYTOX Green uptake. (A) A. niger (○) and P. chrysogenum (•) were incubated with different amounts of AFP in the presence of 0.2 μM SYTOX Green. Fluorescence was measured after 1 h of incubation. Data are averages of triplicate measures. (B) A. niger was incubated with different concentrations of AFP in the presence (•) or absence (○) of 100 mM KCl. Fluorescence was measured as described above. The results of a representative experiment are shown.

Article Snippet: Aspergillus niger ATCC 9029 , 1.

Techniques: Incubation, Fluorescence

Journal:

Article Title: The Antifungal Protein from Aspergillus giganteus Causes Membrane Permeabilization

doi: 10.1128/AAC.47.2.588-593.2003

Figure Lengend Snippet: Localization of AFP within A. niger and P. chrysogenum. (A) FITC-labeled AFP. A. niger (panel A) and P. chrysogenum (panel B) were treated with 10 and 100 μg of FITC-labeled AFP ml, respectively. After 1 h of incubation, the labeled protein was detected by fluorescence microscopy. The control was A. niger treated with 10 μg of FITC-labeled α-sarcin per ml (panel C). Bars, 15 μm. (B) Immunofluorescence. A. niger (panels A and B) and P. chrysogenum (panels C and D) were treated with 10 μg of AFP per ml for 1 h. The protein was detected by immunofluorescence staining with an AFP-specific antibody. As negative controls, P. chrysogenum (panels E and F) and A. niger (panels G and H) were treated with the AFP-specific antibody in the absence of AFP. Panels A, C, E, and G, fluorescence microscopy; panels B, D, F, and H, light-field microscopy. Bars, 15 μm.

Article Snippet: Aspergillus niger ATCC 9029 , 1.

Techniques: Labeling, Incubation, Fluorescence, Microscopy, Control, Immunofluorescence, Staining

Journal: ACS Central Science

Article Title: Dynamic and Functional Profiling of Xylan-Degrading Enzymes in Aspergillus Secretomes Using Activity-Based Probes

doi: 10.1021/acscentsci.9b00221

Figure Lengend Snippet: Fluorescent scans of Cy5 ABP labeled A. niger secretomes induced by xylose or BX. (a) Secretome samples were collected at indicated time points and labeled with the indicated probes. A ∼30 kDa band labeled by 8 was only present in xylan-induced secretomes. (b) Day 6 BX-induced secretomes preincubated with “monosaccharide” competitors, before labeling with 8 . Mono-xylo competitors 1 , 2 , and 4 inhibit labeling of the ∼130 kDa band, suggesting this band corresponds to a β-xylosidase. Glucose configured competitors 22 and 23 have little effect on labeling. (c) BX-induced secretomes preincubated with “disaccharide” competitors, before labeling with 8 . Xylobiose competitors 5 , 6 , 7 , and 9 inhibit labeling of both ∼130 kDa and ∼30 kDa bands, while cellobiose configured 10 shows no effect on labeling. Gel molecular weight markers are given in kilodaltons. Comp. – competitor.

Article Snippet: A. niger strain N402 (a derivative of NRRL3/ATCC 9029/CBS 120.49) was grown in minimal medium containing either 50 mM (0.75% w/v) xylose or 1% w/v beechwood xylan (BX) as the sole carbon source.

Techniques: Labeling, Molecular Weight

Journal: ACS Central Science

Article Title: Dynamic and Functional Profiling of Xylan-Degrading Enzymes in Aspergillus Secretomes Using Activity-Based Probes

doi: 10.1021/acscentsci.9b00221

Figure Lengend Snippet: Peptide signal intensities observed following activity-based protein pulldown from a xylan-induced A. niger secretome. Total MS signal intensity from nonconflicting peptides is shown for (a) XlnD (β-xylosidase), (b) XlnC (GH10 β-xylanase), (c) XlnB (GH11 β-xylanase), (d) BglM (glucosidase), (e) EglA (glucanase), and (f) AxhA (arabinofuranosidase). CAZy GH family names are given next to each enzyme. Box plots show the range of peptide intensities measured in three replicates each of a negative control pulldown with no ABP (DMSO), a β-xylose-configured probe pulldown with ( 2 → 4 ) and without ( 4 ) competitor pretreatment, a β-xylobiose-configured probe pulldown with ( 5 → 9 ) and without ( 9 ) competitor pretreatment, and the total secretome (Total). Full proteomics results can be found in Supplemental File 1 . a.u. – arbitrary units.

Article Snippet: A. niger strain N402 (a derivative of NRRL3/ATCC 9029/CBS 120.49) was grown in minimal medium containing either 50 mM (0.75% w/v) xylose or 1% w/v beechwood xylan (BX) as the sole carbon source.

Techniques: Activity Assay, Negative Control

Journal: ACS Central Science

Article Title: Dynamic and Functional Profiling of Xylan-Degrading Enzymes in Aspergillus Secretomes Using Activity-Based Probes

doi: 10.1021/acscentsci.9b00221

Figure Lengend Snippet: ABP labeling of β-xylosidases and β-xylanases is inhibited by competition with xylanase substrates. ASPACDRAFT_127619 labeling by 8 is inhibited by competition with (a) 4MU-β -d- xylobioside, (b) BX, (c) WAX, and (d) insoluble AZCL-linked WAX. (e) ABP Labeling of XlnC and XlnD in A. niger secretomes is competed by BX. This gel has been contrast adjusted at the dotted line to best show change in labeling intensity for each band. Data points are mean ± standard deviation from three (e) or four (a–d) technical replicates.

Article Snippet: A. niger strain N402 (a derivative of NRRL3/ATCC 9029/CBS 120.49) was grown in minimal medium containing either 50 mM (0.75% w/v) xylose or 1% w/v beechwood xylan (BX) as the sole carbon source.

Techniques: Labeling, Standard Deviation

Journal: ACS Central Science

Article Title: Dynamic and Functional Profiling of Xylan-Degrading Enzymes in Aspergillus Secretomes Using Activity-Based Probes

doi: 10.1021/acscentsci.9b00221

Figure Lengend Snippet: ABP analysis of enzyme thermal denaturation in A. niger secretomes. Both XlnC and XlnD bands are stable up to 60 °C, whereupon XlnC slowly loses activity with increased heating times. XlnC activity was completely abolished within 5 min at 70 °C, whereas extended heating at 80 °C was required to abolish XlnD activity.

Article Snippet: A. niger strain N402 (a derivative of NRRL3/ATCC 9029/CBS 120.49) was grown in minimal medium containing either 50 mM (0.75% w/v) xylose or 1% w/v beechwood xylan (BX) as the sole carbon source.

Techniques: Activity Assay