Heparin binding to human plasma low-density lipoproteins: dependence on heparin sulfation degree and chain length.
Binding between low-density lipoproteins (LDL) and fluorescein-labeled heparin was studied quantitatively with a modified form of a published procedure [Cardin, A. D., Randall, C. I., Hirose, N., & Jackson, R. L. (1987) Biochemistry 26, 5513-5518], using fluorescence anisotropy titrations. Assumption of binding site equivalence satisfactorily interpreted experimental data. Accordingly, the apparent total capacity, n, and the average dissociation constant, Kd, were estimated as n approximately 24 disaccharides per LDL particle and Kd approximately 4 microM in 0.05 M HEPES/0.1 M NaCl, pH 7.4, 22 degrees C. Competition experiments with unlabeled heparins were exploited for the quantitative study of Kd as a function of heparin chain length and sulfation degree (ns = sulfate groups per disaccharide). The former parameter was investigated with a series of bovine lung heparin fractions with Mw ranging from 1,800 to 21,000 and constant sulfation degree (ns = 2.8 +/- 0.1). A series of physically fractionated or chemically modified heparins having 1.2 less than ns less than 3.5 were used to explore the dependence on sulfation degree. LDL affinity was found to increase with increasing both ns and Mw: an empirical Mw-1.6 dependence represented very well the chain length data set; a linear dependence was observed for log Kd as a function of ns, after appropriate allowance was made for chain length differences among samples. This regularity confirmed that LDL-heparin binding is mainly driven by electrostatic forces.(ABSTRACT TRUNCATED AT 250 WORDS)
Gigli
M
M
Farmitalia C. Erba R&D, Erbamont Group, Milan, Italy.
Consonni
A
A
Ghiselli
G
G
Rizzo
V
V
Naggi
A
A
Torri
G
G
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D019793:Fluorescein; D005452:Fluoresceins; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D006493:Heparin / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D013431:Sulfates / Q000378:metabolism*
D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D005452:Fluoresceins; D005456:Fluorescent Dyes; D008077:Lipoproteins, LDL; D013431:Sulfates; D006493:Heparin; D019793:Fluorescein