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Related Concept Videos

Phosphoinositides and PIPs01:42

Phosphoinositides and PIPs

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Phosphoinositides are a group of phospholipids containing a glycerol backbone with two fatty acid chains and a phosphate attached to a myoinositol sugar ring. The inositol head group extends into the cytoplasm, where it is modified by adding phosphate groups to form phosphatidylinositol phosphates or PIPs.
Different phosphoinositides are synthesized and recruited on the cytosolic face of the plasma membrane. The localization of specific phosphoinositides concentrated in separate membrane...
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Related Experiment Video

Updated: Sep 9, 2025

Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry
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Identification of Inositol Phosphate or Phosphoinositide Interacting Proteins by Affinity Chromatography Coupled to Western Blot or Mass Spectrometry

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The Use of Grating-Coupled Interferometry to Study Inositol Pyrophosphate-Protein Interactions.

Kristina Sturm1, Michael Hothorn2

  • 1Structural Plant Biology Laboratory, Department of Plant Sciences, University of Geneva, Geneva, Switzerland.

Methods in Molecular Biology (Clifton, N.J.)
|August 29, 2025
PubMed
Summary

Grating-coupled interferometry (GCI) measures protein-ligand binding kinetics. This study applies GCI with biotinylated inositol pyrophosphates (PP-InsP) to analyze PP-InsP-protein interactions.

Keywords:
Binding kineticsCreoptix WAVEGrating-coupled interferometryInositol pyrophosphatesProtein-ligand interaction

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Preparation of Quality Inositol Pyrophosphates
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Area of Science:

  • Biophysics
  • Biochemistry
  • Molecular Interactions

Background:

  • Grating-coupled interferometry (GCI) is a label-free biophysical technique.
  • Protein-ligand interactions are crucial in cellular signaling and function.
  • Inositol pyrophosphates (PP-InsPs) are signaling molecules with incompletely understood protein interactions.

Purpose of the Study:

  • To apply GCI for characterizing PP-InsP-protein binding kinetics.
  • To demonstrate the utility of biotinylated PP-InsPs as tools for interaction studies.
  • To investigate specific PP-InsP-protein interactions using the Creoptix WAVEsystem.

Main Methods:

  • Utilized grating-coupled interferometry (GCI) on a Creoptix WAVEsystem.
  • Employed biotinylated inositol pyrophosphate (PP-InsP) analogs.
  • Performed kinetic analysis of PP-InsP binding to target proteins.

Main Results:

  • Successfully characterized the binding kinetics of PP-InsP-protein interactions.
  • Demonstrated the effectiveness of GCI for studying these specific molecular interactions.
  • Provided quantitative kinetic data for PP-InsP binding events.

Conclusions:

  • GCI is a powerful method for studying PP-InsP-protein interactions.
  • Biotinylated PP-InsPs are effective reagents for kinetic analysis.
  • This approach advances the understanding of inositol pyrophosphate signaling pathways.