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Related Experiment Video

Updated: Aug 25, 2025

Comparative Proteomic Analysis of Whole Kidney, Medulla, and Cortical Tubules in Diabetic Pathogenesis of Kidney Injury in Mice
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Identification of Potential Megalin/Cubilin Substrates Using Extensive Proteomics Quantification from Kidney

Bei Zhao1, Chengjian Tu1,2, Shichen Shen1,2

  • 1Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214-8033, USA.

The AAPS Journal
|October 17, 2022
PubMed
Summary

Researchers identified potential megalin/cubilin protein substrates by analyzing urine from mice with reduced kidney megalin. This proteomics approach revealed 877 candidate proteins, aiding in understanding kidney reabsorption functions.

Keywords:
Endogenous proteinsKidney reabsorptionLC–MS/MSMegalin/cubilinReceptor-mediated endocytosis

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Area of Science:

  • Nephrology
  • Proteomics
  • Molecular Biology

Background:

  • Megalin and cubilin are key endocytic receptors in kidney proximal tubules responsible for protein reabsorption from filtrate.
  • Understanding their substrates is crucial for elucidating kidney function and potential disease mechanisms.

Purpose of the Study:

  • To identify novel endogenous substrates of megalin and cubilin using a proteomics-based approach.
  • To investigate urinary protein profiles in kidney-specific megalin knockdown (KD) versus wild-type (WT) mice.

Main Methods:

  • Utilized the IonStar proteomics technique to analyze urinary protein content.
  • Compared protein expression in urine from megalin KD mice with that of WT control mice.
  • Quantified protein fold changes and molecular weights to identify potential substrates.

Main Results:

  • Discovered 877 potential megalin/cubilin substrates, including 23 known ones like retinol-binding protein and vitamin D-binding protein.
  • Identified novel substrates with varying molecular weights, with most below 69 kDa.
  • Observed sex differences in substrate numbers potentially linked to kidney megalin expression levels.

Conclusions:

  • Analysis of urine from megalin KD and WT mice is an effective strategy for identifying endogenous megalin/cubilin substrates.
  • The findings provide a foundation for further research into the roles of these proteins in kidney physiology and pathology.