title: High affinity binding between lipoprotein lipase and lipoproteins involves multiple ionic and hydrophobic interactions, does not require enzyme activity, and is modulated by glycosaminoglycans.
abstract: Lipoprotein lipase (LPL) physically associates with lipoproteins and hydrolyzes triglycerides. To characterize the binding of LPL to lipoproteins, we studied the binding of low density lipoproteins (LDL), apolipoprotein (apo) B17, and various apoB-FLAG (DYKDDDDK octapeptide) chimeras to purified LPL. LDL bound to LPL with high affinity (K(d) values of 10(-12) m) similar to that observed for the binding of LDL to its receptors and 1D1, a monoclonal antibody to LDL, and was greater than its affinity for microsomal triglyceride transfer protein. LDL-LPL binding was sensitive to both salt and detergents, indicating the involvement of both hydrophobic and hydrophilic interactions. In contrast, the N-terminal 17% of apoB interacted with LPL mainly via ionic interactions. Binding of various apoB fusion peptides suggested that LPL bound to apoB at multiple sites within apoB17. Tetrahydrolipstatin, a potent enzyme activity inhibitor, had no effect on apoB-LPL binding, indicating that the enzyme activity was not required for apoB binding. LDL-LPL binding was inhibited by monoclonal antibodies that recognize amino acids 380-410 in the C-terminal region of LPL, a region also shown to interact with heparin and LDL receptor-related protein. The LDL-LPL binding was also inhibited by glycosaminoglycans (GAGs); heparin inhibited the interactions by approximately 50% and removal of trace amounts of heparin from LPL preparations increased LDL binding. Thus, we conclude that the high affinity binding between LPL and lipoproteins involves multiple ionic and hydrophobic interactions, does not require enzyme activity and is modulated by GAGs. It is proposed that LPL contains a surface exposed positively charged amino acid cluster that may be important for various physiological interactions of LPL with different biologically important molecules. Moreover, we postulate that by binding to this cluster, GAGs modulate the association between LDL and LPL and the in vivo metabolism of LPL.
authors.affiliation: Department of Biochemistry, School of Medicine, Medical College of Pennsylvania Hahnemann University, Philadelphia, Pennsylvania 19129, USA. mahmoodhussain@netmail.hscbklyn.edu
authors.forename: M M
authors.initials: MM
authors.lastname: Hussain
authors.forename: J C
authors.initials: JC
authors.lastname: Obunike
authors.forename: A
authors.initials: A
authors.lastname: Shaheen
authors.forename: M J
authors.initials: MJ
authors.lastname: Hussain
authors.forename: G S
authors.initials: GS
authors.lastname: Shelness
authors.forename: I J
authors.initials: IJ
authors.lastname: Goldberg
mesh_terms: D000818:Animals; D019556:COS Cells; D002417:Cattle; D004789:Enzyme Activation; D006025:Glycosaminoglycans / Q000737:chemistry* / Q000378:metabolism; D008071:Lipoprotein Lipase / Q000737:chemistry* / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D006025:Glycosaminoglycans; D008074:Lipoproteins; D008071:Lipoprotein Lipase