Sialidase Unmasks Mucin Domain Epitopes of Lubricin.
Lubricin is a secreted, mucin-like glycoprotein and proteoglycan abundant in synovial fluid that provides boundary lubrication and prevents cell adhesion in synovial joints. The antilubricin S6.79 monoclonal antibody recognizes an O-linked glycopeptide epitope in lubricin's mucin domain. The central, long mucin domain of lubricin is extensively O-glycosylated with Gal(β1-3)GalNAc-O-Ser/Thr, and about two thirds of the O-glycosylated sites are capped with sialic acid. Our aim was to determine whether removal of sialic acid by sialidase could improve the detection of lubricin in a number of human tissues using the S6.79 monoclonal antibody. Sialidase treatment caused a dramatic increase in antibody reactivity in human pericardium, splenic capsule and trabeculae, plasma, serum, eye sleep extract, and liver sinusoids. Sialidase had minimal effect on S6.79 antibody reactivity with lubricin in synovial fluid and synovial tissue. These observations suggest that the origin of lubricin in blood may be different from that in synovial fluid and that desialylation of lubricin is essential for unmasking epitopes within the mucin domain.
Department of Biochemistry, Rush University Medical Center, Chicago, Illinois (KAS, TMS).
Kathryn A
KA
Solka
Department of Pathology, Rush University Medical Center, Chicago, Illinois (IJM).
Ira J
IJ
Miller
Department of Biochemistry, Rush University Medical Center, Chicago, Illinois (KAS, TMS).
Thomas M
TM
Schmid
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000939:Epitopes; D004947:Esophagus / Q000737:chemistry; D005143:Eyelids / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D009077:Mucins / Q000737:chemistry* / Q000276:immunology; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D009439:Neuraminidase / Q000737:chemistry*; D009928:Organ Specificity; D010496:Pericardium / Q000737:chemistry; D000072417:Protein Domains; D013154:Spleen / Q000737:chemistry; D013582:Synovial Fluid / Q000737:chemistry; D000099158:Proteoglycan 4
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006023:Glycoproteins; D009077:Mucins; D019158:N-Acetylneuraminic Acid; D009439:Neuraminidase; D000099158:Proteoglycan 4
epitopes; eyelid; liver; lubricin; neuraminidase; pericardium; sialidase; spleen; synovial fluid
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