Journal: Molecular cancer therapeutics

Article Title: Targeting sub-cellular localization through the Polo-Box Domain: non-ATP competitive Inhibitors recapitulate a PLK1 phenotype

doi: 10.1158/1535-7163.MCT-12-0006-T

Figure Lengend Snippet: PLK PBD In vitro binding and cellular activity for FLIP compounds (Ncap-S[pT]PNGL)

Article Snippet: The PLK1 PBD (367–603) and PLK3 PBD (335–646) proteins were obtained from BPS Bioscience Inc. (San Diego, CA) and 250 ng was used per reaction.

Techniques: In Vitro, Binding Assay, Activity Assay

Journal: Frontiers in Molecular Biosciences

Article Title: An Integrated Computational and Experimental Approach to Identifying Inhibitors for SARS-CoV-2 3CL Protease

doi: 10.3389/fmolb.2021.661424

Figure Lengend Snippet: The overview of the proposed workflow to discover new inhibitors of 3CLpro SARS– CoV– 2 .

Article Snippet: In each experimental group, 30 μL of 15 nM purified recombinant 3CLpro (BPS Bioscience Inc., CA) and 10 μL of 500 μM prepared inhibitor solution in 5% aqueous DMSO was added into a black 96-well plate (Nunc U96).

Techniques:

Journal: Frontiers in Molecular Biosciences

Article Title: An Integrated Computational and Experimental Approach to Identifying Inhibitors for SARS-CoV-2 3CL Protease

doi: 10.3389/fmolb.2021.661424

Figure Lengend Snippet: Enzyme activities (A,B) and IC 50 curve (C,D) of newly identified inhibitors, PMPT and CPSQPA. 3CLpro and inhibitors were incubated at 25°C with slow shaking for 2 h, with fluorescence recorded every 3 min (A,B) . The slope of each curve corresponded to the enzyme activity. Relative enzyme activities were calculated as ratio of compound-treated groups and positive control (no inhibitor) to determine IC 50 . Data analysis and curve fitting were conducted using the program Graphpad.

Article Snippet: In each experimental group, 30 μL of 15 nM purified recombinant 3CLpro (BPS Bioscience Inc., CA) and 10 μL of 500 μM prepared inhibitor solution in 5% aqueous DMSO was added into a black 96-well plate (Nunc U96).

Techniques: Incubation, Fluorescence, Activity Assay, Positive Control

Journal: Frontiers in Molecular Biosciences

Article Title: An Integrated Computational and Experimental Approach to Identifying Inhibitors for SARS-CoV-2 3CL Protease

doi: 10.3389/fmolb.2021.661424

Figure Lengend Snippet: Docking conformations of newly identified 3CLpro SARS– CoV– 2 inhibitors: (A,B) PMPT; (C,D) CPSQPA. 3CLpro was modified from the 6LU7 structure by removing the ligand and water. Protein is colored in yellow. Compounds are colored in sky blue. Amino acid residues interacting with the ligands are labeled. Surface of the protein is displayed and colored in gray (B,D) . Hydrogen bonds between PMPT and 3CLpro are shown as black lines (A) . As CPSQPA likely has a net charge of -1 (sulfonamide p K a ’s generally are in the range of 5.0–7.0), the anionic form was docked into the protein (C,D) . S1′, S1, S2, S3, and S4 are the sub-pockets for binding.

Article Snippet: In each experimental group, 30 μL of 15 nM purified recombinant 3CLpro (BPS Bioscience Inc., CA) and 10 μL of 500 μM prepared inhibitor solution in 5% aqueous DMSO was added into a black 96-well plate (Nunc U96).

Techniques: Modification, Labeling, Binding Assay

Journal: Viruses

Article Title: A Novel Hemocyte-Specific Small Protein Participates in White Spot Syndrome Virus Infection via Binding to Viral Envelope Protein

doi: 10.3390/v15010227

Figure Lengend Snippet: Co-location of LvHSSP protein and the hemocytes. rMBP-His represents the results of MBP tag protein, and rMBP-HSSP-His represents the results of LvHSSP recombinant protein. The blue signal of DAPI indicates the nucleus, the green signal of anti-MBP indicates the positive signals of target proteins, the red signal of Di1 indicates the membrane, and the last lanes are the merged picture of previous three signals. Bar = 10 μm.

Article Snippet: After blocking in 5% BSA in TBS at 4 °C overnight, the hemocytes were incubated with mouse anti-MBP antibody (ABclonal, Wuhan, China) in a dilution of 1:500 at room temperature for 1 h, and then incubated with Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Abcam, Shanghai, China) at a dilution of 1:1000 at room temperature for 1 h. Hemocytes were dyed with Di1 (Beyotime, Shanghai, China) for 20 min and DAPI (Beyotime, Shanghai, China) for 5 min before the observation under a laser confocal scanning microscopy (Carl Zeiss LSM 710, Carl Zeiss, Oberkochen, Germany).

Techniques: Recombinant, Membrane