Journal: PLoS ONE

Article Title: Identification of Interactions between Abscisic Acid and Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase

doi: 10.1371/journal.pone.0133033

Figure Lengend Snippet: Raw ITC data of sequential injections of (A) RuBP (0.75 mM) and (B) (+)-ABA (5 mM) titrated against spinach Rubisco (39.42 μM). The processed fit is shown below each binding isotherm. The binding parameters are shown in the inset. The ligands were titrated until saturation was reached showing a specific binding interaction. The data were fitted using the ‘Independent Binding’ model in the NanoAnalyze software and the best fit is represented by the black line.

Article Snippet: A low volume Nano ITC instrument (TA Instruments) with a cell volume of 190 μL was used for this study.

Techniques: Binding Assay, Software

Journal: bioRxiv

Article Title: Unconventional structure and function of PHD domains from Additional Sex Combs-like proteins

doi: 10.1101/2024.12.08.627434

Figure Lengend Snippet: The ASXL PHD domain is unable to bind common histone H3 modifications. (a) ITC for interaction of the ING2 (residues 199–261), ASXL2 (residues 1375–1435), and Drosophila Asx PHD (residues 1610–1668) domains with peptides of H3K4me3 (residues 1–10). Key binding data are listed for the ING2 PHD titration. (b) Fold enrichment of protein signal collected in representative histone H3 ELISA experiments, titrated with samples of the ING2 (residues 199–261), ASXL2 (residues 1375–1435), and Drosophila Asx PHD (residues 1610–1668) domains. Single points correspond to each of the 96 peptides spotted on the ELISA plate, in the order displayed in Appendix Table 1. Values are derived from the fold-enrichment of HRP signal relative to blank. The mean (red) and two standard deviations from the mean (black) are dashed lines. Peptides of H3K4me2 (20 ng) and H3K4me3 (2, 20 ng) are highlighted yellow, orange, and red, respectively (labelled).

Article Snippet: ITC was performed in a low volume TA Affinity ITC instrument (TA instruments).

Techniques: Binding Assay, Titration, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal: PLoS ONE

Article Title: Structural basis of human ORP1-Rab7 interaction for the late-endosome and lysosome targeting

doi: 10.1371/journal.pone.0211724

Figure Lengend Snippet: (A) Binding interface of the ORP1 ANK and Rab7. The residues forming the interface are shown in stick models. The hydrogen bonds are shown in dashed lines. (B) Measurement of binding affinity of ORP1 ANK to Rab7 by isothermal titration calorimetry. The purified Rab7 or Rab7-RILP complex with the concentration of 0.1 mM was titrated with 1 mM of ORP1 ANK. Rab7 was either loaded with GTP or GDP. (C) Overall structure of the ORP1 ANK-Rab7-RILP complex. The structure was constructed by combining the structures of Rab7-RILP (PDB id: 1YHN) and ORP1 -Rab7 (this study) using PyMOL ( https://pymol.org ). The missing C-terminal residues (186–203) of Rab7 were indicated with dotted lines. The C-terminal prenyl groups which anchor Rab7 onto the endosomal membrane are shown in black lines.

Article Snippet: To measure the affinity between ORP1 ANK and Rab7, isothermal titration calorimetry (ITC) was performed using an Affinity ITC calorimeter (low volume cell 190 μl; TA instruments) at 20°C.

Techniques: Binding Assay, Isothermal Titration Calorimetry, Purification, Concentration Assay, Construct, Membrane