Segments in the C-terminal folding domain of lipoprotein lipase important for binding to the low density lipoprotein receptor-related protein and to heparan sulfate proteoglycans

Morten S. Nielsen, Jeanette Brejning, Raquel García, Hanfang Zhang, Michael R. Hayden, Senén Vilaró, Jørgen Gliemann

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Lipoprotein lipase (LpL) can mediate cellular uptake of chylomicron and VLDL remnants via binding to heparan sulfate proteoglycans (HSPG) and the endocytic α2-macroglobulin receptor/low density lipoprotein receptor- related protein (α2MR/LRP). Whereas it is established that the C-terminal folding domain binds to α2MR/LRP, it remains uncertain whether it binds to heparin and to HSPG. To identify segments important for binding to α2MR/LRP and to clarify possible binding to heparin, we produced constructs of the human C-terminal folding domain, LpL-(313-448), and of the fragment LpL- (347-448) in Escherichia coli. In addition to binding to α2MR/LRP, LpL- (313-448) displayed binding to heparin with an affinity similar to that of the LpL monomer, whereas it bound poorly to lipoprotein particles. Moreover, LpL-(313-448) displayed heparin sensitive binding to normal, but not to HSPG deficient Chinese hamster ovary cells. LpL-(313-448) and LpL-(347-448) showed similar affinities for binding to both purified α2MR/LRP and to heparin. Deletion of LpL residues 380-384 abolished the binding to LRP, whereas binding to heparin was unperturbed. The binding to both heparin and α2MR/LRP was essentially abolished following deletion of residues 404-430, and pretreatment of CHO cells with the peptide comprising aa 402-423 inhibited the binding of LpL-(313-448). We conclude that the C-terminal folding domain of human LpL has a site for binding to heparin and to HSPG, presumably involving amino acids within residues 404-430. Two segments of the domain are necessary for efficient binding to α2MR/LRP, one comprising residues 380-384 and another overlapping the segment important for binding to heparin.

Original languageEnglish (US)
Pages (from-to)5821-5827
Number of pages7
JournalJournal of Biological Chemistry
Volume272
Issue number9
DOIs
StatePublished - Feb 28 1997

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LDL-Receptor Related Proteins
Heparan Sulfate Proteoglycans
Lipoprotein Lipase
LDL Receptors
Macroglobulins
Heparin
Proteins
Chylomicron Remnants
Chylomicrons
CHO Cells
Cricetulus
Escherichia coli
Lipoproteins
Ovary
Monomers
Binding Sites

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Segments in the C-terminal folding domain of lipoprotein lipase important for binding to the low density lipoprotein receptor-related protein and to heparan sulfate proteoglycans. / Nielsen, Morten S.; Brejning, Jeanette; García, Raquel; Zhang, Hanfang; Hayden, Michael R.; Vilaró, Senén; Gliemann, Jørgen.

In: Journal of Biological Chemistry, Vol. 272, No. 9, 28.02.1997, p. 5821-5827.

Research output: Contribution to journalArticle

Nielsen, Morten S. ; Brejning, Jeanette ; García, Raquel ; Zhang, Hanfang ; Hayden, Michael R. ; Vilaró, Senén ; Gliemann, Jørgen. / Segments in the C-terminal folding domain of lipoprotein lipase important for binding to the low density lipoprotein receptor-related protein and to heparan sulfate proteoglycans. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 9. pp. 5821-5827.
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abstract = "Lipoprotein lipase (LpL) can mediate cellular uptake of chylomicron and VLDL remnants via binding to heparan sulfate proteoglycans (HSPG) and the endocytic α2-macroglobulin receptor/low density lipoprotein receptor- related protein (α2MR/LRP). Whereas it is established that the C-terminal folding domain binds to α2MR/LRP, it remains uncertain whether it binds to heparin and to HSPG. To identify segments important for binding to α2MR/LRP and to clarify possible binding to heparin, we produced constructs of the human C-terminal folding domain, LpL-(313-448), and of the fragment LpL- (347-448) in Escherichia coli. In addition to binding to α2MR/LRP, LpL- (313-448) displayed binding to heparin with an affinity similar to that of the LpL monomer, whereas it bound poorly to lipoprotein particles. Moreover, LpL-(313-448) displayed heparin sensitive binding to normal, but not to HSPG deficient Chinese hamster ovary cells. LpL-(313-448) and LpL-(347-448) showed similar affinities for binding to both purified α2MR/LRP and to heparin. Deletion of LpL residues 380-384 abolished the binding to LRP, whereas binding to heparin was unperturbed. The binding to both heparin and α2MR/LRP was essentially abolished following deletion of residues 404-430, and pretreatment of CHO cells with the peptide comprising aa 402-423 inhibited the binding of LpL-(313-448). We conclude that the C-terminal folding domain of human LpL has a site for binding to heparin and to HSPG, presumably involving amino acids within residues 404-430. Two segments of the domain are necessary for efficient binding to α2MR/LRP, one comprising residues 380-384 and another overlapping the segment important for binding to heparin.",
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T1 - Segments in the C-terminal folding domain of lipoprotein lipase important for binding to the low density lipoprotein receptor-related protein and to heparan sulfate proteoglycans

AU - Nielsen, Morten S.

AU - Brejning, Jeanette

AU - García, Raquel

AU - Zhang, Hanfang

AU - Hayden, Michael R.

AU - Vilaró, Senén

AU - Gliemann, Jørgen

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