title: Physical-chemical interaction of heparin and human plasma low-density lipoproteins.
abstract: This study characterizes the physical-chemical interactions of heparin with human plasma low-density lipoproteins (LDL). A high reactive heparin (HRH) specific for the surface determinants of LDL was isolated by chromatography of commercial bovine lung heparin on LDL immobilized to AffiGel-10. HRH was derivatized with fluoresceinamine and repurified by affinity chromatography, and its interaction with LDL in solution was monitored by steady-state fluorescence polarization. Binding of LDL to fluoresceinamine-labeled HRH (FL . HRH) was saturable, reversible, and specific; HRH stoichiometrically displaced FL . HRH from the soluble complex, and acetylation of lysine residues on LDL blocked heparin binding. Titration of FL.HRH with excess LDL yielded soluble complexes with two LDL molecules per heparin chain (Mr 13,000) characterized by an apparent Kd of 1 microM. Titration of LDL with excess HRH resulted in two classes of heparin binding with two and five heparin molecules bound per LDL and apparent Kd values of 1 and 10 microM, respectively. At physiological pH and ionic strength, the soluble HRH-LDL complexes were maximally precipitated with 20-50 mM Ca2+. Insoluble complexes contained 2-10 HRH molecules per LDL with the final product stoichiometry dependent on the ratio of the reactants. The affinity of HRH for LDL in the insoluble complexes was estimated between 1 and 10 microM. Insoluble LDL-heparin complexes were readily dissociated with 1.0 M NaCl, and their formation was prevented by acetylation of the lysine residues on LDL.
authors: Merrell Dow Research Institute, Cincinnati, Ohio 45215-6300.
authors: A D
authors: AD
authors: Cardin
authors: C J
authors: CJ
authors: Randall
authors: N
authors: N
authors: Hirose
authors: R L
authors: RL
authors: Jackson
mesh_terms: D000818:Animals; D002417:Cattle; D006493:Heparin / Q000097:blood*; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood*; D008168:Lung; D011485:Protein Binding; D013050:Spectrometry, Fluorescence
chemical_list: D008077:Lipoproteins, LDL; D006493:Heparin