Ultrastructural localization of the C-propeptide released from type II procollagen in fetal bovine growth plate cartilage.
We used immunochemical and immunoelectron gold techniques to determine whether the C-propeptide previously identified in the matrix of endochondral cartilage (CPII) was still a part of the Type 11 procollagen molecule or had been released from it. Guanidinium hydrochloride extraction, followed by SDS-PAGE and Western blotting techniques and immunoelectron localization, revealed that predominantly only the released form (hereafter referred to as released CPII) was detected. The ultrastructural distribution of this CPII was examined with affinity-purified antibodies and with immunogold or immunoperoxidase localization techniques in the presence or absence of embedding resins. These methods yielded similar results. Although no significant amount of this CPII was retained in the matrix after guanidinium hydrochloride extraction, it was present in two recognizable sites under normal conditions, i.e., locally concentrated in a random association with collagen fibrils in the nonmineralized matrix and mainly concentrated in interfibrillar mineralizing sites in the mineralized matrix. These results suggest that the C-propeptide that has been released from Type II procollagen associates with collagen fibrils and then preferentially associates with mineralizing sites when these form in the endochondral cartilage. The significance of this preference for mineral is not known but may have something to do with its high affinity for hydroxyapatite.
Lee
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ER
Joint Diseases Laboratory, Shriners Hospital for Crippled Children, Montreal, Quebec, Canada.
Smith
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CE
Poole
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D000818:Animals; D000906:Antibodies / Q000276:immunology; D002356:Cartilage / Q000196:embryology / Q000378:metabolism* / Q000648:ultrastructure; D002417:Cattle; D005260:Female; D006132:Growth Plate / Q000196:embryology / Q000378:metabolism* / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D010446:Peptide Fragments / Q000378:metabolism*; D011347:Procollagen / Q000378:metabolism*; D011817:Rabbits; D013977:Tibia; D016610:Tissue Embedding
D000906:Antibodies; D010446:Peptide Fragments; D011347:Procollagen; C093212:procollagen type II carboxy-terminal peptide