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Related Experiment Videos

Vesicle-reconstituted low density lipoprotein receptor. Visualization by cryoelectron microscopy.

H Jeon1, G G Shipley

  • 1Departments of Biophysics and Biochemistry, Center for Advanced Biomedical Research, Boston University School of Medicine, Boston, Massachusetts 02118, USA.

The Journal of Biological Chemistry
|July 13, 2000
PubMed
Summary

Researchers visualized the low-density lipoprotein (LDL) receptor using cryoelectron microscopy. This study provides the first low-resolution structural images of the intact LDL receptor, aiding cholesterol homeostasis research.

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Area of Science:

  • Biochemistry
  • Structural Biology
  • Molecular Medicine

Background:

  • The low-density lipoprotein (LDL) receptor is crucial for cellular cholesterol homeostasis.
  • It binds cholesterol-rich lipoproteins via apoB and apoE apoproteins.
  • The receptor is a transmembrane glycoprotein with five identified structural domains.

Purpose of the Study:

  • To obtain the first low-resolution structural images of the intact, full-length bovine LDL receptor.
  • To visualize the receptor in a hydrated state using cryoelectron microscopy (cryoEM).
  • To combine cryoEM with Nanogold labeling for structural insights.

Main Methods:

  • Purification of the LDL receptor from bovine adrenal cortices.
  • Reconstitution of the purified receptor into phosphatidylcholine vesicles.

Related Experiment Videos

  • Imaging of reconstituted LDL receptor using cryoelectron microscopy (cryoEM).
  • Labeling of accessible cysteine residues with Nanogold for structural mapping.

    • Main Results:

    • LDL receptor molecules appeared as elongated, stick-like projections (approx. 120 Å x 45 Å).
    • A short arm (or arms) was observed at the distal end of some projections.
    • CryoEM-Nanogold labeling confirmed the localization of Nanogold to the extracellular domain.

    Conclusions:

    • This study presents the first low-resolution structural images of the reconstituted, full-length bovine LDL receptor.
    • The findings provide a foundation for understanding LDL receptor structure and function in cholesterol transport.
    • CryoEM and Nanogold labeling proved effective for visualizing membrane proteins in a near-native state.