Journal: Scientific Reports

Article Title: Insight into the interaction between the RNA helicase CGH-1 and EDC-3 and its implications

doi: 10.1038/s41598-021-99919-0

Figure Lengend Snippet: The interface of CGH-1 (cyan) and EDC-3 (yellow). ( A ) Interaction details of the Patch 1. Amino acids are shown as side chains. ( B ) Interaction details of the Patch 2. ( C ) Representative curves of ITC assays. ( D ) Effects for the mutations on the binding affinity between CGH-1 RecA2 domain and EDC-3 FDF-FEK peptide.

Article Snippet: In brief, it was carried out on a MicroCal PEAQ-ITC (Malvern) at 20 °C with the following settings: reference power, 5 μcal/s; initial delay, 60 s; stir speed, 750 rpm; spacing time, 120 s. Proteins (or its mutants) and peptides were adjusted to 0.05 mM and 0.5 mM, respectively.

Techniques: Binding Assay

Journal: Scientific Reports

Article Title: Insight into the interaction between the RNA helicase CGH-1 and EDC-3 and its implications

doi: 10.1038/s41598-021-99919-0

Figure Lengend Snippet: Similarity and differences of the binding mode of EDC-3 and CAR-1 for CGH-1. ( A ) The binding mode between CGH-1 RecA2 domain and EDC-3 FDF. Amino acids in the Patch 2 are represented by the side chains. ( B ) The binding mode between CGH-1 RecA2 domain and CAR-1 FDF-TFG. Amino acids in the Patch 3 are represented by the side chains. ( C ) EDC-3 FDF and FEK binding pocket. CGH-1 RecA2 domain is presented as a surface. FEK is marked with blue dotted ellipse. ( D ) CAR-1 FDF and TFG binding pocket. ( E ) Sequence alignment of EDC-3 241–271 and CAR-1 184–214 . The conserved residues SDFDF is marked with a pink box, and FEK is marked with a blue box. Second structures were shown on the top. ( F ) ITC curves: titrating EDC-3 235–271 and CAR-1 184–268 to CGH-1 248–420 , respectively. The ITC data for EDC-3 in Figs. C and 3F are the same data.

Article Snippet: In brief, it was carried out on a MicroCal PEAQ-ITC (Malvern) at 20 °C with the following settings: reference power, 5 μcal/s; initial delay, 60 s; stir speed, 750 rpm; spacing time, 120 s. Proteins (or its mutants) and peptides were adjusted to 0.05 mM and 0.5 mM, respectively.

Techniques: Binding Assay, Sequencing

Journal: Scientific Reports

Article Title: Insight into the interaction between the RNA helicase CGH-1 and EDC-3 and its implications

doi: 10.1038/s41598-021-99919-0

Figure Lengend Snippet: In vitro interaction between CGH-1 and Ce PATR-1 30–67 peptide. ( A ) Representative curves of ITC assays. ( B ) Effects for the CGH-1 mutations on the binding affinity between CGH-1 RecA2 domain and Ce PATR-1 peptide.

Article Snippet: In brief, it was carried out on a MicroCal PEAQ-ITC (Malvern) at 20 °C with the following settings: reference power, 5 μcal/s; initial delay, 60 s; stir speed, 750 rpm; spacing time, 120 s. Proteins (or its mutants) and peptides were adjusted to 0.05 mM and 0.5 mM, respectively.

Techniques: In Vitro, Binding Assay

Journal: Biophysical Reviews

Article Title: Differential scanning fluorimetry followed by microscale thermophoresis and/or isothermal titration calorimetry as an efficient tool for ligand screening

doi: 10.1007/s12551-025-01280-3

Figure Lengend Snippet: Proposed hit screening procedure using DSF followed by ITC/MST methods with main advantages of these approaches and marked further steps for drug design

Article Snippet: Of course, this is always incorporated in commercial software (ITC Data Analysis in Origin ; TA Instruments NanoAnalyze Software Getting Started Guide ) and should be in homemade analysis procedures (SokołOwska et al. ).

Techniques:

Journal: Biophysical Reviews

Article Title: Differential scanning fluorimetry followed by microscale thermophoresis and/or isothermal titration calorimetry as an efficient tool for ligand screening

doi: 10.1007/s12551-025-01280-3

Figure Lengend Snippet: Comparison of most common biophysical method for quantifying ligand binding

Article Snippet: Of course, this is always incorporated in commercial software (ITC Data Analysis in Origin ; TA Instruments NanoAnalyze Software Getting Started Guide ) and should be in homemade analysis procedures (SokołOwska et al. ).

Techniques: Comparison, Binding Assay, High Throughput Screening Assay, Differential Scanning Calorimetry, SPR Assay, Nuclear Magnetic Resonance, Labeling, Mass Spectrometry, Isothermal Titration Calorimetry, Microscale Thermophoresis, Fluorescence, Spectroscopy, Förster Resonance Energy Transfer

Journal: Biophysical Reviews

Article Title: Differential scanning fluorimetry followed by microscale thermophoresis and/or isothermal titration calorimetry as an efficient tool for ligand screening

doi: 10.1007/s12551-025-01280-3

Figure Lengend Snippet: Isothermal titration calorimetry. Standard binding experiment with cartoon representing the bound and unbound protein and two dominating heat effects caused by ligand binding and mixing of components. The heat of each injection gradually reduces as free protein decreases upon ligand binding

Article Snippet: Of course, this is always incorporated in commercial software (ITC Data Analysis in Origin ; TA Instruments NanoAnalyze Software Getting Started Guide ) and should be in homemade analysis procedures (SokołOwska et al. ).

Techniques: Isothermal Titration Calorimetry, Binding Assay, Ligand Binding Assay, Injection

Journal: Biophysical Reviews

Article Title: Differential scanning fluorimetry followed by microscale thermophoresis and/or isothermal titration calorimetry as an efficient tool for ligand screening

doi: 10.1007/s12551-025-01280-3

Figure Lengend Snippet: Comparison of ITC data fitted separately (left panel) and globally (right panel). The example shown is the binding of 4,7-dibromo-5,6-diiodo-1H-benzotriazole to hCK2α, results adapted from (Paprocki et al. ). Black circles show experimental data, grey ones indicate data removed from the analysis, thick blue lines represent the fitted model of two sequential binding sites, and red ones represent the fitted model of a single binding site. The dotted lines boarder the 95% confidence limits for a particular model

Article Snippet: Of course, this is always incorporated in commercial software (ITC Data Analysis in Origin ; TA Instruments NanoAnalyze Software Getting Started Guide ) and should be in homemade analysis procedures (SokołOwska et al. ).

Techniques: Comparison, Binding Assay

Journal: Biophysical Reviews

Article Title: Differential scanning fluorimetry followed by microscale thermophoresis and/or isothermal titration calorimetry as an efficient tool for ligand screening

doi: 10.1007/s12551-025-01280-3

Figure Lengend Snippet: List of studies, in which all three methods: DSF, MST, and ITC were used

Article Snippet: Of course, this is always incorporated in commercial software (ITC Data Analysis in Origin ; TA Instruments NanoAnalyze Software Getting Started Guide ) and should be in homemade analysis procedures (SokołOwska et al. ).

Techniques: Binding Assay, Titration, Kinase Assay