The acidic domain of GPIHBP1 is important for the binding of lipoprotein lipase and chylomicrons.
GPIHBP1, a glycosylphosphatidylinositol-anchored endothelial cell protein of the lymphocyte antigen 6 (Ly6) family, plays a key role in the lipolysis of triglyceride-rich lipoproteins (e.g. chylomicrons). GPIHBP1 is expressed along the luminal surface of endothelial cells of heart, skeletal muscle, and adipose tissue, and GPIHBP1-expressing cells bind lipoprotein lipase (LPL) and chylomicrons avidly. GPIHBP1 contains an amino-terminal acidic domain (amino acids 24-48) that is enriched in aspartate and glutamate residues, and we previously speculated that this domain might be important in binding ligands. To explore the functional importance of the acidic domain, we tested the ability of polyaspartate or polyglutamate peptides to block the binding of ligands to pgsA-745 Chinese hamster ovary cells that overexpress GPIHBP1. Both polyaspartate and polyglutamate blocked LPL and chylomicron binding to GPIHBP1. Also, a rabbit antiserum against the acidic domain of GPIHBP1 blocked LPL and chylomicron binding to GPIHBP1-expressing cells. Replacing the acidic amino acids within GPIHBP1 residues 38-48 with alanine eliminated the ability of GPIHBP1 to bind LPL and chylomicrons. Finally, mutation of the positively charged heparin-binding domains within LPL and apolipoprotein AV abolished the ability of these proteins to bind to GPIHBP1. These studies indicate that the acidic domain of GPIHBP1 is important and that electrostatic interactions play a key role in ligand binding.
Gin
Peter
P
Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, California 90095, USA.
Yin
Liya
L
Davies
Brandon S J
BS
Weinstein
Michael M
MM
Ryan
Robert O
RO
Bensadoun
André
A
Fong
Loren G
LG
Young
Stephen G
SG
Beigneux
Anne P
AP
D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002914:Chylomicrons / Q000737:chemistry*; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008071:Lipoprotein Lipase / Q000737:chemistry / Q000502:physiology*; D051379:Mice; D010455:Peptides / Q000737:chemistry; D011099:Polyglutamic Acid / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D018110:Receptors, Lipoprotein
D002352:Carrier Proteins; D002914:Chylomicrons; C523701:GPIHBP1 protein, human; D008024:Ligands; D010455:Peptides; D018110:Receptors, Lipoprotein; D011099:Polyglutamic Acid; C017645:polyaspartate; D008071:Lipoprotein Lipase