Immunological detection of the EGF-like domain of the core proteins of large proteoglycans from human and baboon cartilage.
Recent data from the literature have shown that cDNA clones for the carboxyterminal domain of the core protein of large proteoglycan monomers from human cartilage contain an EGF-like domain, which appears to undergo alternative splicing. In the present study we have found that articular proteoglycans from human and baboon separated on agarose flat-bed gels and blotted onto nitrocellulose react with a rabbit antiserum to mouse EGF. In addition both forms of the proteoglycans (band I and band II) seen on these gels are reactive. Reactivity is seen with proteoglycans extracted from human articular cartilage of various ages (fetal, newborn, young and aged) and with proteoglycans extracted from cartilage of thanatophoric dysplasia and homozygous achondroplasia. Reactivity is dependent on prior digestion of the nitrocellulose blot with Chase ABC, suggesting masking of epitope by chondroitin sulfate. Reactivity of the EGF antiserum with cartilage proteoglycan core protein was also demonstrated in an ELISA system with core protein as coating antigen. The reactivity appears to reside in a tryptic peptide generated from Chase/keratanase digested core protein. The immunoreactive species migrates as a 68 KDa species on gradient gels. Immunological detection and quantitative analysis of the EGF-like domain could be useful for analysis of various proteoglycan samples.
Stanescu
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V
URA.584-CNRS, Hôpital des Enfants-Malades, Paris, France.
Chaminade
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F
Pham
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TD
D000367:Age Factors; D000368:Aged; D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002358:Cartilage, Articular / Q000276:immunology*; D002648:Child; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D004815:Epidermal Growth Factor / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D007231:Infant, Newborn; D010215:Papio
D000939:Epitopes; D004815:Epidermal Growth Factor