Identification of a protein in squid cranial cartilage with link protein properties

Squid cranial cartilage extracts were found to contain a protein with a molecular mass of 35 kDa immunoreacting with an antiserum against sheep link protein. Because hyaluronan is not detected in this tissue and the structure of proteoglycans is different to that of aggrecan or versican, this observation was studied further. The 35 kDa protein was purified from cartilage extracts and immunolocalised in Western blots by both the polyclonal antibody and the mAb 8A4. It was found that it was able to bind to hyaluronan and to aggrecan. Direct and competitive microplate binding experiments showed that the squid protein binds to G1 domain of aggrecan, similarly to cartilage link protein and, therefore, it could be a link-like protein molecule of squid cranial cartilage. The 35 kDa protein was also able to bind to squid proteoglycan and this suggested that it might participate in squid cartilage proteoglycan aggregate formation.