[Study of the interaction between plasma proteoglycans and LDL by means of fluorescence spectroscopy].
The relevance of the interaction between LDL and PGs in the development of atherosclerotic processes is well known. However, the exact nature of the interaction and the consequent structural and/or conformational modifications of the lipoprotein remain to be clarified. It has been demonstrated that after this interaction the LDL particle is not recognized by specific cellular receptors and enters the scavenger pathway operating in different cell types. These effects have been shown by using aortic PGs, but PGs are also present in the plasma compartment and may interact constantly with LDL, taking part in the regulation of lipid metabolism. In order to assess the capability of plasma PGs to induce LDL modifications, we investigated their interactions by studying the changes in the organizational parameters of LDL by fluorescence spectroscopy. Plasma PGs were isolated by DEAE Sephacel ion exchange chromatography and Sephacryl S300 gel filtration in two different families: a low-charge PG and a high-charge PG. Human LDL was prepared from plasma of normolipemic donors by ultracentrifugal flotation between 1.025-1.045 g/ml. Steady-state anisotropy measures were obtained by analyzing the rotational diffusion rate of DPH after incubation of LDL with plasma PGs in a physiological ratio. In our experimental conditions, LDL incubation with plasma low-charge PG did not modify DPH fluorescence anisotropy, whereas LDL treatment with highly charged PGs induced a marked decrease of this parameter, suggesting a significant effect on LDL microviscosity. The data show that both the charge and the GAG composition of PGs appear to be critical factors in LDL-PG interaction.(ABSTRACT TRUNCATED AT 250 WORDS)
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Istituto di Biologia Applicata, Università di Sassari.
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D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D005454:Fluorescence Polarization; D006025:Glycosaminoglycans / Q000097:blood*; D006801:Humans; D008077:Lipoproteins, LDL / Q000097:blood*; D011485:Protein Binding; D011509:Proteoglycans / Q000097:blood*
D006025:Glycosaminoglycans; D008077:Lipoproteins, LDL; D011509:Proteoglycans
Studio dell'interazione tra proteoglicani plasmatici ed LDL mediante spettroscopia di fluorescenza.