Journal: Journal of medicinal chemistry

Article Title: Protein Stability Effects in Aggregate-Based Enzyme Inhibition

doi: 10.1021/acs.jmedchem.9b01019

Figure Lengend Snippet: Thermodynamic stabilities and binding affinities of TEM-1 mutants. (A) Location of TEM-1 mutations M182T (green), R164S (cyan), G238S (orange), and D179G (red) in TEM-1 M182T mutant PDB structure 1JWP relative to catalytic residues S70, K73, S130, and E166 (pink). (B) Melting temperatures of TEM-1 mutants determined by differential scanning fluorimetry (DSF) (see Table 1 for Tm and ΔΔG values). (C) The competitive displacement of 100 nM 5-maleimido-fluorescein-ribosomal protein L2 globular domain (5-MF-L2gd) bound to 20 μM Sor/CR colloids by TEM-1 stability mutants.

Article Snippet: 35 , 36 Incubation was performed for 20 nM TEM-1 and its mutants with varying concentrations of Sor/CR, Fulvestrant, TIPT, and Miconazole (MedChemExpress, HY-10201; Millipore Sigma, C6277; MedChemExpress, HY-13636; Spectrum Chemical, T0126; Santa Cruz Biotechnology, sc-204806) for 5 min at room temperature in 50 mM KPi, pH 7.0.

Techniques: Binding Assay, Mutagenesis

Journal: Journal of medicinal chemistry

Article Title: Protein Stability Effects in Aggregate-Based Enzyme Inhibition

doi: 10.1021/acs.jmedchem.9b01019

Figure Lengend Snippet: Thermodynamic stabilities and binding affinities of TEM-1 mutants in complex with moxalactam. (A) The chemical structure of moxalactam. (B) Change in melting temperatures of TEM-1 mutants incubated with 100-fold molar excess of β-lactamase inhibitor moxalactam relative to respective apo-enzymes, determined by DSF (see Table 2). The competitive displacement of 100 nM 5-MF-L2gd bound to 20 μM Sor/CR colloids by TEM-1 stability mutants, (C) WT and WT-inhibitor complex, (D) R164S and R164S-inhibitor complex, (E) D179G and D179G-inhibitor complex, and (F) M182T and M182T-inhibitor complex (see Tables 1 and ​and22 for EC50 values).

Article Snippet: 35 , 36 Incubation was performed for 20 nM TEM-1 and its mutants with varying concentrations of Sor/CR, Fulvestrant, TIPT, and Miconazole (MedChemExpress, HY-10201; Millipore Sigma, C6277; MedChemExpress, HY-13636; Spectrum Chemical, T0126; Santa Cruz Biotechnology, sc-204806) for 5 min at room temperature in 50 mM KPi, pH 7.0.

Techniques: Binding Assay, Incubation

Journal: Journal of medicinal chemistry

Article Title: Protein Stability Effects in Aggregate-Based Enzyme Inhibition

doi: 10.1021/acs.jmedchem.9b01019

Figure Lengend Snippet: TEM-1 inhibition dose–response curves against (A) Sor/CR, (B) Fulvestrant, (C) TIPT, and (D) Miconazole colloids (see Table 3 for IC50 values).

Article Snippet: 35 , 36 Incubation was performed for 20 nM TEM-1 and its mutants with varying concentrations of Sor/CR, Fulvestrant, TIPT, and Miconazole (MedChemExpress, HY-10201; Millipore Sigma, C6277; MedChemExpress, HY-13636; Spectrum Chemical, T0126; Santa Cruz Biotechnology, sc-204806) for 5 min at room temperature in 50 mM KPi, pH 7.0.

Techniques: Inhibition

Journal: ACS Omega

Article Title: Phage Display-Derived Peptide for the Specific Binding of SARS-CoV-2

doi: 10.1021/acsomega.1c04873

Figure Lengend Snippet: Peptide 1 can specifically bind to the SARS-CoV-2 virus. (a) Schematic of the experimental procedure for the characterization of peptide binding with the heat-inactivated SARS-CoV-2 virus using the magnetic bead assay and TEM. (b–e) Biotinylated peptide 1 was immobilized on SA-coated magnetic beads to capture the virus. The gold nanoparticle-labeled SARS-CoV-2 antibody was used to detect the virus. The viruses and gold nanoparticles were visualized using TEM. The influenza virus served as a negative control. (b,c) Four individual magnetic beads with the addition of 0.67 and 0.067 mg/mL of the SARS-CoV-2 target to the surface, showing that gold nanoparticles (dashed red circle) have conjugated onto the magnetic beads. (d,e) Four individual magnetic beads with the addition of 0.67 and 0.067 mg/mL of influenza virus. The scale bar is 100 nm.

Article Snippet: The SARS envelope protein antibody used in the TEM assay was purchased from Novus Biologicals, LLC., Centennial, CO. N-Hydroxysuccinimide (NHS)-activated gold nanoparticles with a diameter of 40 nm were purchased from Cytodiagnostics Inc., Canada.

Techniques: Virus, Binding Assay, Magnetic Beads, Labeling, Negative Control