Journal: PLoS ONE

Article Title: In-Cell Protease Assay Systems Based on Trans-Localizing Molecular Beacon Proteins Using HCV Protease as a Model System

doi: 10.1371/journal.pone.0059710

Figure Lengend Snippet: Before cleavage, GFP was found in the plasma membrane (ring shape, before cleavage). Cleavage of the type I MBR by HCV NS3 protease resulted in the translocation of GFP to the cytosol (dispersed image, after cleavage).

Article Snippet: A known inhibitor of the HCV NS3 protease, Ac-Asp-D-Gla-Leu-Ile-b-cyclohexyl-Ala-Cyc-OH (N-1725.0001, Bachem, Germany) was purchased and dissolved in dimethyl sulfoxide.

Techniques: Clinical Proteomics, Membrane, Translocation Assay

Journal: PLoS ONE

Article Title: In-Cell Protease Assay Systems Based on Trans-Localizing Molecular Beacon Proteins Using HCV Protease as a Model System

doi: 10.1371/journal.pone.0059710

Figure Lengend Snippet: Type II MBRs were constructed to contain a mitochondrial targeting sequence, MSRA as a masked translocation signal. The masking protein used was GAPDH or GFP for mono- or dual-color type II MBRs, respectively. A . Cleavage of mono-color type II MBR by HCV NS3 protease activates the mitochondrial translocation signal of MSRA, resulting in translocation of GFP from the cytosol (before cleavage) to the mitochondria (after cleavage). B . Cleavage of dual-color type II MBR which contains both GFP and RFP, results in translocation of the RFP from the cytosol (before cleavage) to mitochondria while GFP remains in the cytosol (after cleavage) following cleavage. GFP; green fluorescent protein, RFP; red fluorescent protein.

Article Snippet: A known inhibitor of the HCV NS3 protease, Ac-Asp-D-Gla-Leu-Ile-b-cyclohexyl-Ala-Cyc-OH (N-1725.0001, Bachem, Germany) was purchased and dissolved in dimethyl sulfoxide.

Techniques: Construct, Sequencing, Translocation Assay

Journal: PLoS ONE

Article Title: In-Cell Protease Assay Systems Based on Trans-Localizing Molecular Beacon Proteins Using HCV Protease as a Model System

doi: 10.1371/journal.pone.0059710

Figure Lengend Snippet: The cleavage of type I MBR ( A ) and type II MBR ( B ) by HCV NS3 protease was monitored in the presence of a known peptide derivative inhibitor, Ac-Asp-D-Gla-Leu-Ile-β-cyclohexyl-Ala-Cyc-OH. Both type I and type II MBRs exhibited significant dose-response at nanomolar concentrations.

Article Snippet: A known inhibitor of the HCV NS3 protease, Ac-Asp-D-Gla-Leu-Ile-b-cyclohexyl-Ala-Cyc-OH (N-1725.0001, Bachem, Germany) was purchased and dissolved in dimethyl sulfoxide.

Techniques:

Journal: PLoS ONE

Article Title: In-Cell Protease Assay Systems Based on Trans-Localizing Molecular Beacon Proteins Using HCV Protease as a Model System

doi: 10.1371/journal.pone.0059710

Figure Lengend Snippet: An HCV NS3 protease inhibitory RNA aptamer was prepared and tested. Cells co-transfected with vectors encoding type I MBR and HCV NS3 protease were and treated with the RNA aptamer. The inhibition ratio was determined from the number of cells exhibiting a dispersed pattern relative to the total number of cells examined.

Article Snippet: A known inhibitor of the HCV NS3 protease, Ac-Asp-D-Gla-Leu-Ile-b-cyclohexyl-Ala-Cyc-OH (N-1725.0001, Bachem, Germany) was purchased and dissolved in dimethyl sulfoxide.

Techniques: Transfection, Inhibition