Journal: Marine Drugs

Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer

doi: 10.3390/md15090290

Figure Lengend Snippet: Dose-response curves for kempopeptins B and C ( 2 and 3 ) against serine proteases. ( A ) Trypsin; ( B ) plasmin; and ( C ) matriptase. The dose-response is presented as % fold inhibition against solvent control (DMSO). Data are presented as the mean ± SD, n = 3.

Article Snippet: Recombinant human matriptase (R&D Systems; 3946-SE-010) and recombinant human desmoglein 2 FC Chimera (R&D Systems; 947-DM-100) or recombinant protein of human CUB domain containing protein 1 (CDCP1), transcript variant 1 (OriGene; TP320633, Rockville, MD, USA) were incubated in an enzyme to substrate ratio (1:5) for 30 min, 1 h, 2 h or 3 h, pH 9.0, at 37 °C.

Techniques: Inhibition, Solvent, Control

Journal: Marine Drugs

Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer

doi: 10.3390/md15090290

Figure Lengend Snippet: IC 50 values of kempopeptins B ( 2 ) and C ( 3 ) against serine proteases.

Article Snippet: Recombinant human matriptase (R&D Systems; 3946-SE-010) and recombinant human desmoglein 2 FC Chimera (R&D Systems; 947-DM-100) or recombinant protein of human CUB domain containing protein 1 (CDCP1), transcript variant 1 (OriGene; TP320633, Rockville, MD, USA) were incubated in an enzyme to substrate ratio (1:5) for 30 min, 1 h, 2 h or 3 h, pH 9.0, at 37 °C.

Techniques:

Journal: Marine Drugs

Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer

doi: 10.3390/md15090290

Figure Lengend Snippet: Docked structures of kempopeptin B (pink) and kempopeptin C (green) in matriptase. ( A ) The atomic sizes of the Br (red) and Cl (green) are displayed as spheres; ( B ) the residues of the matriptase enzyme in close proximity to Br (Trp) and Cl (Phe and Thr) are displayed in stick representation (cyan).

Article Snippet: Recombinant human matriptase (R&D Systems; 3946-SE-010) and recombinant human desmoglein 2 FC Chimera (R&D Systems; 947-DM-100) or recombinant protein of human CUB domain containing protein 1 (CDCP1), transcript variant 1 (OriGene; TP320633, Rockville, MD, USA) were incubated in an enzyme to substrate ratio (1:5) for 30 min, 1 h, 2 h or 3 h, pH 9.0, at 37 °C.

Techniques:

Journal: Marine Drugs

Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer

doi: 10.3390/md15090290

Figure Lengend Snippet: The effect of kempopeptin C ( 3 ) on the in vitro cleavage of CDCP1 and desmoglein-2 (Dsg-2) by matriptase. ( A , B ) Incubation of recombinant human CDCP1 with recombinant human matriptase (enzyme to substrate ratio 1:5) in 50 mM Tris, 50 mM NaCl, 0.01% Tween 20 (pH 9.0) at 37 °C for 30 min and 2 h in the presence and absence of different concentrations (10‒0.01 µM) of kempopeptin C ( 3 ). ( C ) Incubation of recombinant human Dsg-2 with recombinant human matriptase (enzyme to substrate ratio 1:5) in 50 mM Tris, 50 mM NaCl, 0.01% tween 20 (pH 9.0) at 37 °C for 3 h in the presence and absence of different concentrations (10‒0.01 µM) of kempopeptin C ( 3 ). Fragments were separated on SDS-PAGE under reducing conditions and silver stained. ( D ) Densitometric analysis of Dsg-2 bands in the SDS-PAGE in C.

Article Snippet: Recombinant human matriptase (R&D Systems; 3946-SE-010) and recombinant human desmoglein 2 FC Chimera (R&D Systems; 947-DM-100) or recombinant protein of human CUB domain containing protein 1 (CDCP1), transcript variant 1 (OriGene; TP320633, Rockville, MD, USA) were incubated in an enzyme to substrate ratio (1:5) for 30 min, 1 h, 2 h or 3 h, pH 9.0, at 37 °C.

Techniques: In Vitro, Incubation, Recombinant, SDS Page, Staining

Journal: Marine Drugs

Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer

doi: 10.3390/md15090290

Figure Lengend Snippet: The effect of kempopeptins B and C ( 2 and 3 ) on CDCP1 cleavage in MDA-MB-231. ( A ) MDA-MB-231 cells were treated for 72 h with different concentrations of the test compounds. Following 72 h, the lysates were collected and analyzed by Western blot. ( B ) MDA-MB-231 cells were treated at different time points with 100 µM of kempopeptin C ( 3 ). Following 6 h, 12 h and 24 h, the lysates were collected and analyzed by Western blot. Dexamethasone (Dex) was used as a positive control. ( C ) MDA-MB-231 cells treated for 24 h with different concentrations of 3 in the presence of 10 nM recombinant human matriptase. Cell lysates were harvested and analyzed by Western blot. ( D ) Densitometric analysis of the full-length, LMW of CDCP1 and actin bands in (C).

Article Snippet: Recombinant human matriptase (R&D Systems; 3946-SE-010) and recombinant human desmoglein 2 FC Chimera (R&D Systems; 947-DM-100) or recombinant protein of human CUB domain containing protein 1 (CDCP1), transcript variant 1 (OriGene; TP320633, Rockville, MD, USA) were incubated in an enzyme to substrate ratio (1:5) for 30 min, 1 h, 2 h or 3 h, pH 9.0, at 37 °C.

Techniques: Western Blot, Positive Control, Recombinant