Journal: Methods in enzymology

Article Title: A High-Throughput Biological Calorimetry Core – Steps to Startup, Run, and Maintain a Multi-user Facility

doi: 10.1016/bs.mie.2015.07.024

Figure Lengend Snippet: VP-cap DSC Instrument from MicroCal. This instrument allows up to six 96 well trays to be loaded and programmed without user intervention. Figure used with permission from Malvern.

Article Snippet: A MicroCal Auto-iTC200 ITC instrument and a Microcal VP-cap DSC instrument were purchased from General Electric (Microcal wing of GE, which is now with Malvern instruments) each of which has 96-well formats.

Techniques:

Journal: bioRxiv

Article Title: The structure of HiSiaQM defines the architecture of tripartite ATP-independent periplasmic (TRAP) transporters

doi: 10.1101/2021.12.03.471092

Figure Lengend Snippet: Isothermal titration calorimetry experiment between HiSiaP and the HiSiaP-VHH (left) and binding curve (middle). The thermodynamic parameters from the experiment are shown on the right.

Article Snippet: ITC experiments were performed as described before in Peter et al. on a MicroCal PEAQ device from Malvern Panalytical, using corresponding MicroCalPEAQ-ITC software (version 1.21) for experiment design, measurement and analysis.

Techniques: Isothermal Titration Calorimetry, Binding Assay

Journal: bioRxiv

Article Title: The structure of HiSiaQM defines the architecture of tripartite ATP-independent periplasmic (TRAP) transporters

doi: 10.1101/2021.12.03.471092

Figure Lengend Snippet: SEC elution profiles of the HiSiaP wildtype and mutants that showed a growth defect in the complementation assay. The SEC runs were performed using the standard overexpression and purification protocol as described in the methods. Eluted fractions were checked via SDS-PAGE, shown below the corresponding chromatograms. The binding of the HiSiaP mutants to sialic acid was analyzed with isothermal titration calorimetry experiments. An exemplary differential power graph and a binding curve is shown for each mutant. The mean binding parameters from three measurements for each mutant are specified.

Article Snippet: ITC experiments were performed as described before in Peter et al. on a MicroCal PEAQ device from Malvern Panalytical, using corresponding MicroCalPEAQ-ITC software (version 1.21) for experiment design, measurement and analysis.

Techniques: Over Expression, Purification, SDS Page, Binding Assay, Isothermal Titration Calorimetry, Mutagenesis

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