Collagenase-extractable proteoglycans from lesion-free areas of human aorta.
Different proteoglycan (PG) populations were isolated from normal human aorta by extraction of minced tissue with 4M GuHCl and by further digestion of the residue with collagenase. Dissociative extraction induced a complete disappearance of Alcian Blue positive material, which was demonstrable by transmission electron microscopy before the treatment around collagen fibrils and in pericellular areas. However, 4M GuHCl extraction solubilized only an average of 60% of aorta total hexuronate content. Collagenase treatment of the residue resulted in a complete loss of collagen fibril organization, which was coupled with a further hexuronate recovery, accounting for about one third of total tissue content. The bulk of PGs obtained in collagenase digest was retained by Sepharose CL-4B column. Their sulphated glycosaminoglycan (GAG) composition differed from PGs extracted with 4M GuHCl, containing only chondroitin sulphate (CS) and heparan sulphate (HS), without detectable traces of dermatan sulphate (DS). Moreover, they contained hyaluronic acid. The results obtained by agarose polyacrylamide gel electrophoresis (APGE) and Octyl-Sepharose chromatography, followed by further APGE and Sepharose CL-4B gel-filtration, carried out before and after treatment with Chondroitinase ABC and AC and Heparinase I and III, suggested that collagenase digest contained different PG populations, carrying mainly either CS or HS chains. Moreover, HS containing PGs showed higher hydrodynamic size and stronger properties of hydrophobic interactions than CS containing PGs.
Bardoni
A
A
Dipartimento di Biochimica, Università di Pavia.
Salvini
R
R
Pallavicini
G
G
Passi
A
A
Rindi
S
S
Quacci
D
D
Dall'Orbo
C
C
De Luca
G
G
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001013:Aorta, Thoracic / Q000737:chemistry* / Q000473:pathology; D002850:Chromatography, Gel; D003013:Clostridium / Q000201:enzymology; D017364:Collagenases; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D011489:Protein Denaturation; D011509:Proteoglycans / Q000032:analysis / Q000302:isolation & purification* / Q000502:physiology
D006146:Guanidines; D011509:Proteoglycans; D017364:Collagenases; D019791:Guanidine