Fibrillin: evidence that chondroitin sulphate proteoglycans are components of microfibrils and associate with newly synthesised monomers.
We have investigated the potential association of proteoglycans with intact fibrillin-containing microfibrils from foetal bovine elastic tissues and with newly synthesised fibrillin in human and bovine cell cultures. Microfibril integrity was disrupted by chondroitinase ABC lyase and chondroitinase AC lyase, but not by keratanase or hyaluronidase. Following chondroitinase treatment, beads were disrupted but the underlying fibrillar scaffold appeared intact. Cuprolinic blue was prominently associated with beaded domains at a critical electrolyte concentration. Electron-dense rods were often associated with cuprolinic blue-treated microfibrils isolated from fixed tissues. Positive staining revealed charged foci at the beads. Newly synthesised fibrillin could be labelled with 35S TransLabel, [3H]glucosamine or 35SO4 but its electrophoretic mobility was not influenced by treatment with chondroitinase ABC or AC lyase. A diffuse 35SO4-labelled chondroitinase-sensitive component with a resistant band (Mr 35000) co-immunoprecipitated with fibrillin. These experiments indicate that chondroitin sulphate proteoglycans associate with fibrillin and contribute to microfibril assembly. This association has major implications for microfibril function in health and disease.
Kielty
C M
CM
Department of Medicine, University of Manchester, UK. cay.kielty@man.ac.uk
Whittaker
S P
SP
Shuttleworth
C A
CA
D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D001011:Aorta; D002417:Cattle; D002478:Cells, Cultured; D002808:Chondroitin Lyases / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D000071837:Fibrillins; D006026:Glycoside Hydrolases; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D008840:Microfilament Proteins / Q000096:biosynthesis / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D011233:Precipitin Tests; D001616:beta-Galactosidase / Q000378:metabolism
D011508:Chondroitin Sulfate Proteoglycans; D000071837:Fibrillins; D008840:Microfilament Proteins; D006026:Glycoside Hydrolases; C026093:keratan-sulfate endo-1,4-beta-galactosidase; D001616:beta-Galactosidase; D006821:Hyaluronoglucosaminidase; D002808:Chondroitin Lyases