Isolation and characterization of low sulfated heparan sulfate sequences with affinity for lipoprotein lipase.
Lipoprotein lipase (LPL), which is an important enzyme in lipid metabolism, binds to heparan sulfate (HS) proteoglycans. This interaction is crucial for several aspects of LPL function, such as intracellular/extracellular transport and high capacity attachment to cell surfaces. Retention of LPL on the capillary walls, and elsewhere, via HS chains is most likely affected by the quality and quantity of HS present. Earlier studies have demonstrated that LPL interacts with highly sulfated HS and heparin oligosaccharides. Since such structures are relatively rare in endothelial HS, we have re-addressed the question of physiological ligand structures for LPL by affinity purification of end-labeled oligosaccharides originating from heparin and HS on immobilized LPL. By a combination of chemical modification and fragmentation of the bound material we identified that the bound fraction contained modestly sulfated oligosaccharides with an average sulfation of one O-sulfate per disaccharide unit and tolerates N-acetylated glucosamine residues. Therefore LPL, containing several clusters of positive charges on each subunit, may constitute an ideal structure for a protein that needs to bind with reasonable affinity to a variety of modestly sulfated sequences of the type that is abundant in HS chains.
Spillmann
Dorothe
D
Department of Medical Biochemistry and Microbiology, University of Uppsala, SE-751 23 Uppsala, Sweden. Dorothe.Spillmann@imbim.uu.se
Lookene
Aivar
A
Olivecrona
Gunilla
G
D000107:Acetylation; D000818:Animals; D001667:Binding, Competitive; D002240:Carbohydrate Sequence; D002417:Cattle; D004730:Endothelium, Vascular / Q000737:chemistry / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006497:Heparan Sulfate / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006868:Hydrolysis; D008071:Lipoprotein Lipase / Q000737:chemistry / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013552:Swine
D005944:Glucosamine; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D008071:Lipoprotein Lipase; D009844:Oligosaccharides