Formation of heparan sulfate or chondroitin/dermatan sulfate on recombinant domain I of mouse perlecan expressed in Chinese hamster ovary cells.
Recombinant domain I of mouse perlecan was expressed in Chinese hamster ovary (CHO K1) cells and affinity purified on Ni-agarose. Gel chromatography followed by characterization of glycosaminoglycans by the use of glycosaminoglycan lyases showed that the recombinant proteoglycans contained, on average, three glycosaminoglycan chains of heparan sulfate or chondroitin/dermatan sulfate of approximately 12 kDa median size. These data demonstrate that domain I has functional sites for attachment of glycosaminoglycans and indicate that the glycosaminoglycan chains of native perlecan are grouped at its N-terminal end. This, in turn, suggests that the likely function of domain I in perlecan would be to provide for the addition of glycosaminoglycan chains to the core protein.
Kokenyesi
R
R
Connective Tissue Research Laboratory, Edith Nourse Rogers Memorial Veterans Hospital, Bedford, MA 01730, USA.
Silbert
J E
JE
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002809:Chondroitin Sulfates / Q000096:biosynthesis*; D002846:Chromatography, Affinity; D006224:Cricetinae; D017931:DNA Primers; D003871:Dermatan Sulfate / Q000096:biosynthesis*; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000302:isolation & purification; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011509:Proteoglycans / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism*; D014162:Transfection
D002809:Chondroitin Sulfates; D017931:DNA Primers; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans; D011994:Recombinant Proteins; C071927:perlecan