Journal: Cell Chemical Biology

Article Title: Development of potent and selective inhibitors targeting the papain-like protease of SARS-CoV-2

doi: 10.1016/j.chembiol.2021.04.020

Figure Lengend Snippet: Inhibitor 19 is a potent and selective SCoV2 PLpro inhibitor (A) Chemical structures of 12 and 19 . (B and C) Inhibitor 19 was docked into the binding pocket of 12 in SCoV2 PLpro (chain A, PDB: 7E35 ). Superimposition of 19 (purple sticks) and 12 (azure sticks) indicated additional interactions between 19 and Tyr269-Gln 270 (green sticks in B) of SCoV2 PLpro (surface in C). (D) SPR analysis of 19 binding to SCoV2 PLpro; GRL0617 was used as a control. K d and SD were calculated based on three independent experiments. (E) A bar graph showing the inhibition of 11 DUBs or DUB-like proteases by 19 (10 μM) (n = 3). Data are presented as the mean ± SD (n = 3).

Article Snippet: SPR analysis , GE Healthcare , Cat# BR-1005-30.

Techniques: Binding Assay, Inhibition

Journal: Cell Chemical Biology

Article Title: Development of potent and selective inhibitors targeting the papain-like protease of SARS-CoV-2

doi: 10.1016/j.chembiol.2021.04.020

Figure Lengend Snippet:

Article Snippet: SPR analysis , GE Healthcare , Cat# BR-1005-30.

Techniques: Produced, Recombinant, Activity Assay, Luciferase, Stability Assay, Software

Journal: The Journal of Biological Chemistry

Article Title: Tyrosine phosphorylation switching of a G protein

doi: 10.1074/jbc.RA117.000163

Figure Lengend Snippet: AtGPA1 Y166E changes its binding specificity with AtRGS1. A, direct interaction between GST-tagged AtRGS1 (RGS + Ct) and His-tagged AtGPA1 wildtype or Y166E mutant in the presence of GDP, GTPγS, or GDP-AlF4− was detected by an in vitro pulldown assay. Protein complexes were purified by glutathione-Sepharose, separated by SDS-PAGE, and detected by anti-GST or anti-His antibodies. B, the intensities of His-AtGPA1 wildtype and Y166E mutant were quantitated with ImageJ and normalized as relative values to each interaction in the presence of GDP-AlF4−. The quantitative results were expressed as the means ± S.D. (error bars) of four experiments. Statistical significance was determined by ANOVA. *, differences with p values of <0.05. **, differences with p values of <0.01. The binding affinity constants of AtRGS1 and AtGPA1 wildtype or Y166E mutant were measured by surface plasmon resonance with a ProteOn XPR36 instrument. The GST-tagged AtRGS1 (RGS + Ct) was immobilized on the surface of a GLC sensor chip as ligand, and the His-tagged AtGPA1 wildtype or Y166E in the GDP-bound, GTPγS-bound, or GDP-AlF4−-bound state were diluted into dosage concentrations as analyte. The affinity constants were calculated by kinetic analysis (C–H) or equilibrium analysis (I-K) and are shown in Table 2. IP, immunoprecipitation; IB, immunoblotting.

Article Snippet: Surface plasmon resonance Binding affinity constants were measured by the ProteOn XPR36 protein interaction array system and a ProteOn TM GLC Sensor Chip (Bio-Rad, catalog no. 1765011).

Techniques: Binding Assay, Mutagenesis, In Vitro, Purification, SDS Page, SPR Assay, Immunoprecipitation, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Tyrosine phosphorylation switching of a G protein

doi: 10.1074/jbc.RA117.000163

Figure Lengend Snippet: Summary of affinity constants between AtRGS1 and Gα subunits Binding affinity constants were measured using a ProteOn XPR36 surface plasmon resonance instrument. For the kinetic analysis, the original curves were fit with a 1:1 Langmuir binding model. k a , association rate constant; k d , dissociation rate constant; K D , calculated by k a / k d ; R max , maximum response; χ 2 , the average of squared residuals. For the equilibrium analysis, K D was calculated by response units at steady state. RU, response units; NA, no specific binding was detected with the GDP-bound state of wildtype AtGPA1.

Article Snippet: Surface plasmon resonance Binding affinity constants were measured by the ProteOn XPR36 protein interaction array system and a ProteOn TM GLC Sensor Chip (Bio-Rad, catalog no. 1765011).

Techniques: Binding Assay, SPR Assay

Journal: Structure (London, England : 1993)

Article Title: Structural adaptation in its orphan domain engenders betaglycan with an alternate mode of growth factor binding relative to endoglin

doi: 10.1016/j.str.2019.06.010

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: CM-5 SPR sensor chip , GE Lifesciences , Cat# 29149604.

Techniques: Recombinant, Software