Mutagenesis in four candidate heparin binding regions (residues 279-282, 291-304, 390-393, and 439-448) and identification of residues affecting heparin binding of human lipoprotein lipase

Y. Ma, H. E. Henderson, M. S. Liu, H. Zhang, I. J. Forsythe, I. Clarke-Lewis, M. R. Hayden, J. D. Brunzell

Research output: Contribution to journalArticle

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Abstract

Lipoprotein lipase (LPL) interaction with membrane-associated polyanions is a critical component of normal catalytic function. Two strong candidate binding regions, rich in arginine and lysine residues, have been defined in the N-terminal domain (aa279-282 and aa292-304) that show homology to the heparin-binding consensus sequences -X-B-B-X-B-X- and -X-B-B-B-X-X-B-X-, respectively. Additional candidate regions appear in the C-terminal domain, (residues 390-393), which are homologous to the thrombospondin heparin- binding repeat, and the positively charged terminal decapeptide (residues 439-448). To determine residues and domains critical to heparin binding, we have generated different LPL mutants that have alanine substitutions of single arginine and lysine residues and sequence interchanges with the homologous hepatic (HL) and pancreatic (PL) lipases. The mutant cDNAs were expressed in COS-1 cells and catalytically active mutants were assessed for binding to heparin Sepharose. All the alanine substitutions within the two regions homologous to the heparin-binding consensus sequences in the N- terminal domain either abolished activity or produced a lowering of heparin binding affinity. None of the mutants in the C-terminal domain of LPL showed a loss of activity or a reduction in heparin binding affinity. These data demonstrate that charged residues at positions 279-282 and 292-304 of LPL are important for heparin binding affinity whereas the residues 390-393 and 439- 448 in the C-terminal domain are not involved in heparin binding.

Original languageEnglish (US)
Pages (from-to)2049-2059
Number of pages11
JournalJournal of Lipid Research
Volume35
Issue number11
StatePublished - Jan 1 1994

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Mutagenesis
Heparin
Lipoprotein Lipase
Consensus Sequence
Alanine
Lysine
Arginine
Substitution reactions
Thrombospondins
human LPL protein
COS Cells
Interchanges
Lipase
Complementary DNA
Membranes
Liver

Keywords

  • heparin binding
  • in vitro mutagenesis
  • mutations

ASJC Scopus subject areas

  • Endocrinology

Cite this

Mutagenesis in four candidate heparin binding regions (residues 279-282, 291-304, 390-393, and 439-448) and identification of residues affecting heparin binding of human lipoprotein lipase. / Ma, Y.; Henderson, H. E.; Liu, M. S.; Zhang, H.; Forsythe, I. J.; Clarke-Lewis, I.; Hayden, M. R.; Brunzell, J. D.

In: Journal of Lipid Research, Vol. 35, No. 11, 01.01.1994, p. 2049-2059.

Research output: Contribution to journalArticle

Ma, Y. ; Henderson, H. E. ; Liu, M. S. ; Zhang, H. ; Forsythe, I. J. ; Clarke-Lewis, I. ; Hayden, M. R. ; Brunzell, J. D. / Mutagenesis in four candidate heparin binding regions (residues 279-282, 291-304, 390-393, and 439-448) and identification of residues affecting heparin binding of human lipoprotein lipase. In: Journal of Lipid Research. 1994 ; Vol. 35, No. 11. pp. 2049-2059.
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AU - Liu, M. S.

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AU - Hayden, M. R.

AU - Brunzell, J. D.

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