Cloning and primary structure of neurocan, a developmentally regulated, aggregating chondroitin sulfate proteoglycan of brain.

摘要: We have obtained the complete coding sequence of neurocan, a chondroitin sulfate proteoglycan rat brain which is developmentally regulated with respect to its molecular size, concentration, carbohydrate composition, sulfation, and immunocytochemical localization. Two degenerate oligonucleotides, based on amino acid data from isolated adult by immunoaffinity chromatography 1D1 monoclonal antibody, were used as sense antisense primers in polymerase chain reaction cDNA library template generate an unambiguous probe. A second probe for N-terminal portion early postnatal form was reverse transcription/polymerase reaction. The composite overlapping clones 5.2-kilobases (kb) long, including 1.3 kb 3‘-untranslated 76 base pairs 5‘-untranslated sequence. An open reading frame 1257 acids encodes protein mass 136 kDa containing 10 peptide sequences present and/or proteoglycans. deduced revealed 22-amino signal followed immunoglobulin domain, tandem repeats characteristic hyaluronic acid-binding region aggregating proteoglycans, RGDS C-terminal (amino 951-1215) has approximately 60% identity regions C termini fibroblast cartilage versican aggrecan, two epidermal growth factor-like domains, lectin-like complement regulatory protein-like central 595-amino neurocan no homology other reported sequences. contains six potential N-glycosylation sites 25 threonine O-glycosylation sites. In (which represents half neurocan) single 32-kDa 4-sulfate linked at serin-944, whereas three additional attachment (only are utilized) larger species. corresponding having or aggrecan hybridized band 7.5 Northern blots mRNA both 4-day (but not muscle, kidney, liver, lung mRNA), indicating that brain, 68-kDa core protein, generated vivo proteolytic processing 136-kDa species predominant brain.(ABSTRACT TRUNCATED AT 400 WORDS)