Arterial lumican. Properties of a corneal-type keratan sulfate proteoglycan from bovine aorta.
A glycoprotein reactive with antibodies against corneal keratan sulfate proteoglycan (KSPG) was purified 300-fold from extracts of bovine aorta using DEAE ion-exchange, gel-filtration, hydrophobic interaction, and reverse-phase chromatographic separations. The intact glycoprotein was 70-80 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Deglycosylation with endo-beta-galactosidase and N-glycanase reduced the size to 48 and 37 kDa, respectively, similar to the large isoforms of corneal KSPG. N-terminal amino acid sequence of the arterial KSPG was identical with lumican, the 37B isoform of corneal KSPG, and the arterial KSPG reacted with an antibody to synthetic peptide duplicating this sequence. Arterial KSPG and corneal lumican displayed identical tryptic maps. Arterial lumican contains fucose and mannose in amounts similar to corneal KSPG, but galactose, glucosamine, and sulfate were reduced compared to KSPG from cornea. Treatment of arterial lumican with endo-beta-galactosidase released 8-9 mol of glucosamine and galactose per mol of protein as oligosaccharides. These eluted as neutral, nonsulfated oligosaccharides on high pH anion-exchange chromatography. The size of arterial lumican was not altered by glycosidases having specificity for sulfated keratan sulfate, nor was the charge of the lumican molecule altered by digestion with endo-beta-galactosidase. These data show arterial lumican to be a glycoprotein containing unsulfated lactosaminoglycan chains. Abundance of low sulfate lumican in many tissues indicates that this protein occurs predominantly as a glycoprotein rather than as the more widely studied, highly sulfated proteoglycan present in the cornea.
Funderburgh
J L
JL
Division of Biology, Kansas State University, Manhattan 66506.
Funderburgh
M L
ML
Mann
M M
MM
Conrad
G W
GW
D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000737:chemistry*; D002417:Cattle; D011508:Chondroitin Sulfate Proteoglycans / Q000235:genetics / Q000302:isolation & purification*; D002853:Chromatography, Liquid; D003315:Cornea / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007632:Keratan Sulfate / Q000235:genetics / Q000302:isolation & purification*; D000071234:Lumican; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D014357:Trypsin
D011508:Chondroitin Sulfate Proteoglycans; D000071234:Lumican; D007632:Keratan Sulfate; D006026:Glycoside Hydrolases; D014357:Trypsin