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Identification of the calcitonin receptor by chemical cross‐linking and photoaffinity labeling in human cancer cell lines
Author(s) -
Moseley J. M.,
Smith P.,
Martin T. J.
Publication year - 1986
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1002/jbmr.5650010308
Subject(s) - calcitonin , calcitonin receptor , cell culture , receptor , chemistry , biochemistry , calcitonin gene related peptide , biology , endocrinology , neuropeptide , genetics
Two methods have been used to covalently cross‐link [ 125 I]‐salmon calcitonin to its receptor on a human lung carcinoma cell line, BEN, and the human breast cancer cell lines T47D and MCF 7. The first method was to use a specific photoaffinity derivative of salmon calcitonin and the second employed the chemical cross‐linker, disuccinimidyl suberate. In both cases a cross‐linked component of approximate molecular weight 80–90,000 on BEN cells was identified by polyacrylamide gel electrophoresis. This is consistent with the size of the cross‐linked component found on T47D breast cancer cells using the photoactive salmon calcitonin as described in previous work. Disuccinimidyl suberate was unable to cross‐link [ 125 I]‐salmon calcitonin either on T47D or MCF cells. However, photoactive salmon calcitonin cross‐linked to a component of approximately 80–90,000 M r on the MCF 7 cells. Thus, whereas the photoactive salmon calcitonin could cross‐link a similar receptor component in all cell lines, the ability of disuccinimidyl suberate to do so was apparently cell specific. These data confirm that the calcitonin receptor comprises a component of approximately 85,000 M r in cell lines examined thus far.