PROTEOGLYCANS IN MACROPHAGES - CHARACTERIZATION AND POSSIBLE ROLE IN THE CELLULAR UPTAKE OF LIPOPROTEINS

Citation
B. Halvorsen et al., PROTEOGLYCANS IN MACROPHAGES - CHARACTERIZATION AND POSSIBLE ROLE IN THE CELLULAR UPTAKE OF LIPOPROTEINS, Biochemical journal, 331, 1998, pp. 743-752
Citations number
53
Categorie Soggetti
Biology
Journal title
ISSN journal
02646021
Volume
331
Year of publication
1998
Part
3
Pages
743 - 752
Database
ISI
SICI code
0264-6021(1998)331:<743:PIM-CA>2.0.ZU;2-Z
Abstract
The murine macrophage cell line J774 was incubated with [S-35]sulphate . The cell-associated S-35-labelled macromolecules were shown to be pr oteoglycans and glycosaminoglycans in similar amounts. The possible pr esence of cell-surface proteoglycans was investigated by incubating [S -35]sulphate-labelled cells with trypsin for 15 min. The released mate rial contained approx. 70 % free glycosaminoglycan chains and 30 % pro teoglycans. The latter component was demonstrated by HNO2 treatment to contain heparan sulphate. In the total cell fraction not treated with trypsin a small but significant portion was shown to be chondroitin s ulphate proteoglycan, The cell-associated glycosaminoglycans contained both chondroitin sulphate and heparan sulphate. To investigate possib le biological functions of cell-surface proteoglycans in macrophages, cells were incubated with NaClO3 to inhibit sulphation of proteoglycan s and beta-D-xyloside to abrogate proteoglycan expression. The uptake of oxidized I-125-tyraminylcellobiose-labelled low-density lipoprotein (I-125-TC-LDL) was typically two to three times higher than that of n ative I-125-TC-LDL in untreated J774 cells. The cellular uptake at 37 degrees C of native I-125-TC-LDL was decreased 25% after both NaClO3 a nd xyloside treatment, whereas the uptake of oxidized I-125-TC-LDL was decreased 35 % after both types of treatment. The mRNA levels for the scavenger receptor A-II and the LDL receptor were not affected by NaC lO3 or xyloside treatment, Furthermore, fluid-phase endocytosis, measu red as uptake of horseradish peroxidase, and receptor-mediated endocyt osis, measured as uptake of I-125-TC-ovalbumin, were not affected by N aClO3 treatment of J774 cells. Removal of cell-surface chondroitin sul phate with chondroitinase ABC decreased only the binding of native I-1 25-TC-LDL, whereas removal of heparan sulphate with heparitinase decre ased the binding of both oxidized and native I-125-TC-LDL. Addition of lipoprotein lipase increased the uptake of oxidized I-125-TC-LDL 1.7 times and the uptake of native I-125-TC-LDL 2.1 times. The binding of the former was more sensitive to NaClO3 treatment than the latter. The results presented support the notion that some of the uptake pathways for lipoproteins in the foam-cell-forming macrophages depend on the p resence of cell-surface heparan sulphate and chondroitin sulphate.