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Related Experiment Video

Updated: May 1, 2026

Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes
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Inositol phosphates induce DAPI fluorescence shift.

Bernadett Kolozsvari1, Federica Parisi1, Adolfo Saiardi1

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Summary

Inositol phosphates like IP6 and IP5 bind to DAPI, altering its fluorescence. This discovery enables rapid monitoring of inositol phosphates and enzymatic reactions in plant seeds.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Polyphosphate (polyP) and inositol phosphates share biophysical similarities due to their phosphate groups.
  • Polyphosphate is known to modify the excitation-emission spectra of DAPI.

Purpose of the Study:

  • To investigate if inositol phosphates also alter DAPI fluorescence.
  • To explore the application of this phenomenon in studying inositol phosphate metabolism and quantification.

Main Methods:

  • Complexing DAPI with various inositol phosphates (IP6, IP5, IP3, IP4).
  • Measuring excitation-emission spectra of DAPI-inositol phosphate complexes.
  • Utilizing DAPI fluorescence shifts to monitor enzymatic activity (inositol polyphosphate multikinase, phytase phosphatase).
  • Quantifying inositol hexakisphosphate (IP6) in plant seeds using a DAPI-IP6 fluorescence standard curve.

Main Results:

  • DAPI-IP6 and DAPI-IP5 complexes exhibit fluorescence emission around 550 nm upon excitation at 410-420 nm.
  • IP3 and IP4 do not induce a significant shift in DAPI fluorescence.
  • The DAPI fluorescence shift was successfully used to study enzymatic reactions.
  • Borlotti beans showed the highest IP6 content (9.4 mg/g dry mass), while Indian urad beans had the lowest (3.2 mg/g dry mass).

Conclusions:

  • Inositol phosphates IP6 and IP5 interact with DAPI, causing a detectable fluorescence shift.
  • This DAPI-based assay provides a rapid and reliable method for monitoring inositol phosphate levels and enzymatic conversions.
  • The method is applicable for quantifying IP6 in plant seeds, with potential for broader applications in biological research.