C cell (parafollicular cell) -- immunoreactive thyroglobulin: purification, identification and immunological characterization.
In relation to our earlier finding that the thyroglobulin-like material responsible for the cytochemical immunoreaction of C cells was obtained in the peak I fraction of Bio-Gel A-5 m, which included faster sedimenting components of thyroglobulin, the present study has identified the positive reacting component and clarified its immunochemical and immunohistochemical properties. 1. The peak I fraction of dog and hog thyroglobulin was chromatographed on a Bio-Gel A-50 m column. Antiserum to the faster eluted peak I'1 only immunoreacted with C cells. The peak I'1 was then refiltered on Bio-Gel A-150 m column. Antiserum to peak I''1 fraction of both species which was eluted in the first part had high immune specificity for C cells. 2. When 4-30% and 2-16% continuous gradient gels of polyacrylamide were employed, peak I''1 represented a single electrophoretic band corresponding to the component with the largest molecular weight in thyroglobulin. The protein was named C-thyroglobulin. The molecular weight was approximately 2,600,000, four times as large as 19 S, as calculated by relative mobility on the 2-16% gradient gel. 3. In double diffusion tests, anti-peak I''1 antiserum produced two immunoprecipitin lines with its own antigen. The reaction was different from that of anti-19 S antiserum which formed a single line. 4. On immunoperoxidase staining, anti-peak I''1 antiserum reacted to C cells in exactly the same way as anti-calcitonin antiserum. 5. When anti-peak I''1 antiserum was absorbed with calcitonin, the subsequent reaction of the C cells was greatly decreased. The absorption of anti-calcitonin antiserum with increased amounts of peak I''1 abolished the C cell reaction. On the basis of these observations, the possibility that C-thyroglobulin is a biosynthetic precursor of calcitonin exists.
D000818:Animals; D002116:Calcitonin / Q000276:immunology; D004285:Dogs; D000939:Epitopes; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D008970:Molecular Weight; D013552:Swine; D013954:Thyroglobulin / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D013961:Thyroid Gland / Q000032:analysis* / Q000166:cytology
D000939:Epitopes; D002116:Calcitonin; D013954:Thyroglobulin