N-Glycosylation regulates endothelial lipase-mediated phospholipid hydrolysis in apoE- and apoA-I-containing high density lipoproteins

Skropeta, D; Settasatian, C; McMahon, MR; Shearston, K; Caiazza, D; McGrath, KC; Jin, W; Rader, DJ; Barter, PJ; Rye, KA

N-Glycosylation regulates endothelial lipase-mediated phospholipid hydrolysis in apoE- and apoA-I-containing high density lipoproteins

Skropeta, D Settasatian, C McMahon, MR Shearston, K Caiazza, D McGrath, KC

Jin, W Rader, DJ Barter, PJ Rye, KA

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Publication Type:: Journal Article
Citation:: Journal of Lipid Research, 2007, 48 (9), pp. 2047 - 2057
Issue Date:: 2007-09-01

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Field	Value	Language
dc.contributor.author	Skropeta, D	en_US
dc.contributor.author	Settasatian, C	en_US
dc.contributor.author	McMahon, MR	en_US
dc.contributor.author	Shearston, K	en_US
dc.contributor.author	Caiazza, D	en_US
dc.contributor.author	McGrath, KC https://orcid.org/0000-0002-6244-3929	en_US
dc.contributor.author	Jin, W	en_US
dc.contributor.author	Rader, DJ	en_US
dc.contributor.author	Barter, PJ	en_US
dc.contributor.author	Rye, KA	en_US
dc.date.issued	2007-09-01	en_US
dc.identifier.citation	Journal of Lipid Research, 2007, 48 (9), pp. 2047 - 2057	en_US
dc.identifier.issn	0022-2275	en_US
dc.identifier.uri	http://hdl.handle.net/10453/14910
dc.description.abstract	Endothelial lipase (EL) is a member of the triglyceride lipase gene family with high phospholipase and low triacylglycerol lipase activities and a distinct preference for hydrolyzing phospholipids in HDL. EL has five potential N-glycosylation sites, four of which are glycosylated. The aim of this study was to determine how glycosylation affects the phospholipase activity of EL in physiologically relevant substrates. Site-directed mutants of EL were generated by replacing asparagine (N) 62, 118, 375, and 473 with alanine (A). These glycan-deficient mutants were used to investigate the kinetics of phospholipid hydrolysis in fully characterized preparations of spherical reconstituted high density lipoprotein (rHDL) containing apolipoprotein E2 (apoE2) [(E2)rHDL], apoE3 [(E3)rHDL], apoE4 [(E4)rHDL], or apoA-I [(A-I)rHDL] as the sole apolipoprotein. Wild-type EL hydrolyzed the phospholipids in (A-I)rHDL, (E2)rHDL, (E3)rHDL, and (E4)rHDL to similar extents. The phospholipase activities of EL N118A, EL N375A, and EL N473A were significantly diminished relative to that of wild-type EL, with the greatest reduction being apparent for (E3)rHDL. The phospholipase activity of EL N62A was increased up to 6-fold relative to that of wild-type EL, with the greatest enhancement of activity being observed for (E2)rHDL. These data show that individual N-linked glycans have unique and important effects on the phospholipase activity and substrate specificity of EL. Copyright © 2007 by the American Society for Biochemistry and Molecular Biology, Inc.	en_US
dc.relation.ispartof	Journal of Lipid Research	en_US
dc.relation.isbasedon	10.1194/jlr.M700248-JLR200	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Cells, Cultured	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Lipase	en_US
dc.subject.mesh	Lipoproteins, HDL	en_US
dc.subject.mesh	Phospholipids	en_US
dc.subject.mesh	Apolipoprotein A-I	en_US
dc.subject.mesh	Apolipoproteins E	en_US
dc.subject.mesh	Recombinant Proteins	en_US
dc.subject.mesh	Amino Acid Substitution	en_US
dc.subject.mesh	Glycosylation	en_US
dc.subject.mesh	Hydrolysis	en_US
dc.subject.mesh	Kinetics	en_US
dc.title	N-Glycosylation regulates endothelial lipase-mediated phospholipid hydrolysis in apoE- and apoA-I-containing high density lipoproteins	en_US
dc.type	Journal Article
utslib.citation.volume	9	en_US
utslib.citation.volume	48	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	1101 Medical Biochemistry and Metabolomics	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
utslib.copyright.status	closed_access
pubs.issue	9	en_US
pubs.publication-status	Published	en_US
pubs.volume	48	en_US

Abstract:

Endothelial lipase (EL) is a member of the triglyceride lipase gene family with high phospholipase and low triacylglycerol lipase activities and a distinct preference for hydrolyzing phospholipids in HDL. EL has five potential N-glycosylation sites, four of which are glycosylated. The aim of this study was to determine how glycosylation affects the phospholipase activity of EL in physiologically relevant substrates. Site-directed mutants of EL were generated by replacing asparagine (N) 62, 118, 375, and 473 with alanine (A). These glycan-deficient mutants were used to investigate the kinetics of phospholipid hydrolysis in fully characterized preparations of spherical reconstituted high density lipoprotein (rHDL) containing apolipoprotein E2 (apoE2) [(E2)rHDL], apoE3 [(E3)rHDL], apoE4 [(E4)rHDL], or apoA-I [(A-I)rHDL] as the sole apolipoprotein. Wild-type EL hydrolyzed the phospholipids in (A-I)rHDL, (E2)rHDL, (E3)rHDL, and (E4)rHDL to similar extents. The phospholipase activities of EL N118A, EL N375A, and EL N473A were significantly diminished relative to that of wild-type EL, with the greatest reduction being apparent for (E3)rHDL. The phospholipase activity of EL N62A was increased up to 6-fold relative to that of wild-type EL, with the greatest enhancement of activity being observed for (E2)rHDL. These data show that individual N-linked glycans have unique and important effects on the phospholipase activity and substrate specificity of EL. Copyright © 2007 by the American Society for Biochemistry and Molecular Biology, Inc.

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http://hdl.handle.net/10453/14910