Cellular localization and characterization of proteins that bind high density lipoprotein

B. Hokland, Armando J Mendez, J. F. Oram

Research output: Contribution to journalArticle

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Abstract

High density lipoprotein (HDL) stimulates excretion of excess intracellular cholesterol from cells, presumably by interacting with a cell-surface receptor. A 110 kDa membrane protein that is a candidate for the HDL receptor has been identified by ligand blot analysis. In this study we determined the cellular localization of this and other HDL- binding proteins and characterized their properties. The plasma membranes (PM) of cultured bovine aortic endothelial cells were labeled with trace amounts of [3H]cholesterol, and cell homogenates were fractionated on sucrose and Percoll gradients. Ligand blot analysis of homogenates of cultured bovine aortic endothelial cells demonstrated that cells contain multiple proteins that bind HDL3, including a major membrane protein with an apparent M(r) of 110 kDa and two minor ones with M(r) of 105 and 130 kDa. The gradient distribution of the 105, 110, and 130 kDa HDL- binding proteins mirrored that of labeled cholesterol and 5'- nucleotidase, both PM markers. Treatment of intact cells with the water- soluble cross-linker bis(sulfosuccinimidyl)suberate abolished the HDL binding activity of the 110 and 130 kDa proteins but not that of the 105 kDa protein. These findings suggest that the 105, 110, and 130 kDa HDL- binding proteins are localized to the PM and that at least two of these proteins are exposed to the extracellular fluid. Solubilized 110 and 130 kDa proteins were retained on wheat-germ agglutinin and abrin lectin columns, showing that they are glycoproteins. The cellular localization and physical properties of the 110 and 130 kDa proteins suggest that they may play a role in binding of HDL to the cell surface.

Original languageEnglish
Pages (from-to)1335-1342
Number of pages8
JournalJournal of Lipid Research
Volume33
Issue number9
StatePublished - Jan 1 1992
Externally publishedYes

Fingerprint

HDL Lipoproteins
Cell membranes
Proteins
Cholesterol
Endothelial cells
Cell Membrane
Abrin
Membrane Proteins
Endothelial Cells
Ligands
5'-Nucleotidase
Wheat Germ Agglutinins
Extracellular Fluid
Cell Surface Receptors
Lectins
Sucrose
Glycoproteins
Physical properties
Cells
Fluids

Keywords

  • cellular high density lipoprotein-binding proteins

ASJC Scopus subject areas

  • Endocrinology

Cite this

Cellular localization and characterization of proteins that bind high density lipoprotein. / Hokland, B.; Mendez, Armando J; Oram, J. F.

In: Journal of Lipid Research, Vol. 33, No. 9, 01.01.1992, p. 1335-1342.

Research output: Contribution to journalArticle

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