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Related Experiment Videos

Endothelial cells synthesize and process apolipoprotein B

P Sivaram1, T Vanni-Reyes, I J Goldberg

  • 1Division of Preventive Medicine and Nutrition, Department of Medicine, Columbia University, New York, New York 10032, USA.

The Journal of Biological Chemistry
|June 21, 1996
PubMed
Summary

Endothelial cells synthesize apolipoprotein B (apoB) and a 116-kDa fragment. This fragment, derived from full-length apoB, binds lipoprotein lipase (LPL), suggesting a role in lipid metabolism.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Endothelial cells possess a lipoprotein lipase (LPL)-binding protein with sequence homology to apolipoprotein (apo) B.
  • Previous research indicated a potential link between endothelial cells and apoB.

Purpose of the Study:

  • To investigate whether endothelial cells synthesize apolipoprotein B (apoB) mRNA and protein.
  • To characterize the apoB protein produced by endothelial cells and its potential fragments.

Main Methods:

  • Reverse transcription polymerase chain reaction (RT-PCR) was used to detect apoB mRNA in bovine and human endothelial cells.
  • Metabolic labeling with radioactive isotopes followed by immunoprecipitation with anti-apoB antibodies was employed to identify apoB protein.
  • Ligand blot assays were performed to assess the LPL-binding activity of apoB fragments.

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Main Results:

  • Endothelial cells express both full-length apoB mRNA and protein (approximately 550 kDa).
  • A 116-kDa apoB fragment was identified, appearing as full-length apoB degraded and exhibiting LPL-binding capacity.
  • Heparin treatment released the 116-kDa fragment, and oleic acid did not significantly alter apoB production.

Conclusions:

  • Endothelial cells actively synthesize apolipoprotein B (apoB).
  • A 116-kDa apoB fragment is generated through cleavage of full-length apoB.
  • This 116-kDa fragment functions as a lipoprotein lipase (LPL)-binding protein, implicating endothelial apoB in lipid transport and metabolism.