Boundary lubrication by lubricin is mediated by O-linked beta(1-3)Gal-GalNAc oligosaccharides.
Lubrication of mammalian joints is mediated by lubricin, a product of megakaryocyte stimulating factor gene (MSF; GenBank accession #U70136) expression. Lubricin (M(r) approximately 240 kDa) is a mucinous glycoprotein which is 50% (w/w) post-translationally modified with beta(1-3)Gal-GalNAc incompletely capped with NeuAc, and lubricates apposed cartilaginous surfaces in the boundary mode through an unknown mechanism. Both bovine and human lubricin were purified from synovial fluid and digested with recombinant glycosidases. Released oligosaccharides were identified and quantified by fluorophore assisted carbohydrate electrophoresis (FACE). Corresponding digests of human lubricin were also assayed in a friction apparatus oscillating latex rubber against polished glass at a pressure of 0.35 x 10(6) N/m(2) and the coefficient of friction (mu) was measured. Digestion with alpha2,3-neuraminidase decreased lubricating ability by 19.3%. Partial removal of beta(1-3)Gal-GalNAc moieties by endo-alpha-N-acetyl-D-galactosaminidase reduced lubricating ability by 77.2%. Human lubricin digested with combined alpha2,3-neuraminidase and beta1-3,6-galactosidase continued to lubricate at 52.2% of its nominal value. Both bovine and human lubricin released 48.6% and 54.4% of total beta(1-3)Gal-GalNAc sidechains following digestion with endo-alpha-N-acetyl-D-galactosaminidase. Biological boundary lubrication by synovial fluid in vitro is provided primarily by extensive O-linked beta(1-3)Gal-GalNAc.
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Department of Medicine, Section of Emergency Medicine, Brown University School of Medicine, Providence, RI 02903, USA. gregory_jay_MD@brown.edu
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D000116:Acetylgalactosamine / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D000818:Animals; D002417:Cattle; D004586:Electrophoresis / Q000379:methods; D005456:Fluorescent Dyes; D005690:Galactose / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000502:physiology; D006031:Glycosylation; D006596:Hexosaminidases / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008153:Lubrication; D009844:Oligosaccharides / Q000737:chemistry* / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D013583:Synovial Membrane / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D000099158:Proteoglycan 4
D000116:Acetylgalactosamine; D005456:Fluorescent Dyes; D005690:Galactose; D006023:Glycoproteins; D006596:Hexosaminidases; D009844:Oligosaccharides; D011994:Recombinant Proteins; D000099158:Proteoglycan 4