Assessing the Role of the Glycosylphosphatidylinositol-anchored High Density Lipoprotein-binding Protein 1 (GPIHBP1) Three-finger Domain in Binding Lipoprotein Lipase

Anne P Beigneux; Brandon S. J Davies; Shelly Tat; Jenny Chen; Peter Gin; Constance V Voss; Michael M Weinstein; André Bensadoun; Clive R Pullinger; Loren G Fong; Stephen G Young

doi:10.1074/jbc.M111.242024

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Assessing the Role of the Glycosylphosphatidylinositol-anchored High Density Lipoprotein-binding Protein 1 (GPIHBP1) Three-finger Domain in Binding Lipoprotein Lipase

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Assessing the Role of the Glycosylphosphatidylinositol-anchored High Density Lipoprotein-binding Protein 1 (GPIHBP1) Three-finger Domain in Binding Lipoprotein Lipase

Anne P Beigneux, Brandon S. J Davies, Shelly Tat, Jenny Chen, Peter Gin, Constance V Voss, Michael M Weinstein, André Bensadoun, Clive R Pullinger, Loren G Fong, …

The Journal of biological chemistry, Vol.286(22), pp.19735-19743

06/03/2011

DOI: 10.1074/jbc.M111.242024

PMCID: PMC3103352

PMID: 21478160

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Abstract

Glycosylphosphatidylinositol-anchored high density lipoprotein-binding protein 1 (GPIHBP1) is an endothelial cell protein that transports lipoprotein lipase (LPL) from the subendothelial spaces to the capillary lumen. GPIHBP1 contains two main structural motifs, an amino-terminal acidic domain enriched in aspartates and glutamates and a lymphocyte antigen 6 (Ly6) motif containing 10 cysteines. All of the cysteines in the Ly6 domain are disulfide-bonded, causing the protein to assume a three-fingered structure. The acidic domain of GPIHBP1 is known to be important for LPL binding, but the involvement of the Ly6 domain in LPL binding requires further study. To assess the importance of the Ly6 domain, we created a series of GPIHBP1 mutants in which each residue of the Ly6 domain was changed to alanine. The mutant proteins were expressed in Chinese hamster ovary (CHO) cells, and their expression level on the cell surface and their ability to bind LPL were assessed with an immunofluorescence microscopy assay and a Western blot assay. We identified 12 amino acids within GPIHBP1, aside from the conserved cysteines, that are important for LPL binding; nine of those were clustered in finger 2 of the GPIHBP1 three-fingered motif. The defective GPIHBP1 proteins also lacked the ability to transport LPL from the basolateral to the apical surface of endothelial cells. Our studies demonstrate that the Ly6 domain of GPIHBP1 is important for the ability of GPIHBP1 to bind and transport LPL.

Lipoprotein Metabolism

LPL

Metabolism

Lipase

Lipolysis

Ly6 Domain

Endothelium

Protein-Protein Interactions

GPIHBP1

Details

Title: Subtitle: Assessing the Role of the Glycosylphosphatidylinositol-anchored High Density Lipoprotein-binding Protein 1 (GPIHBP1) Three-finger Domain in Binding Lipoprotein Lipase
Creators: Anne P Beigneux - From the Departments of
Brandon S. J Davies - From the Departments of
Shelly Tat - From the Departments of
Jenny Chen - Human Genetics, David Geffen School of Medicine, UCLA, Los Angeles, California 90095
Peter Gin - From the Departments of
Constance V Voss - From the Departments of
Michael M Weinstein - From the Departments of
André Bensadoun - the
Clive R Pullinger - the
Loren G Fong - From the Departments of
Stephen G Young - From the Departments of
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.286(22), pp.19735-19743
DOI: 10.1074/jbc.M111.242024
PMID: 21478160
PMCID: PMC3103352
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology; 9650 Rockville Pike, Bethesda, MD 20814, U.S.A
Grant note: R01 HL094732; P01 HL090553; R01 HL087228 / National Institutes of Health
Alternative title: GPIHBP1 LPL-binding Domain
Language: English
Date published: 06/03/2011
Academic Unit: Fraternal Order of Eagles Diabetes Research Center; Biochemistry and Molecular Biology
Record Identifier: 9984025395302771

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