Moseley J M, Smith P, Martin T J
University of Melbourne, Department of Medicine, Repatriation General Hospital, Heidelberg, Victoria, Australia.
J Bone Miner Res. 1986 Jun;1(3):293-7. doi: 10.1002/jbmr.5650010308.
Two methods have been used to covalently cross-link [125I]-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 [125I]-salmon calcitonin either on T47D or MCF cells. However, photoactive salmon calcitonin cross-linked to a component of approximately 80-90,000 Mr 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 Mr in cell lines examined thus far.
已采用两种方法将[125I] - 鲑鱼降钙素与人类肺癌细胞系BEN、人类乳腺癌细胞系T47D和MCF 7上的受体进行共价交联。第一种方法是使用鲑鱼降钙素的特异性光亲和衍生物，第二种方法则使用化学交联剂辛二酸二琥珀酰亚胺酯。在这两种情况下，通过聚丙烯酰胺凝胶电泳在BEN细胞上鉴定出了一种分子量约为80 - 90,000的交联成分。这与先前工作中使用光活性鲑鱼降钙素在T47D乳腺癌细胞上发现的交联成分大小一致。辛二酸二琥珀酰亚胺酯无法在T47D或MCF细胞上使[125I] - 鲑鱼降钙素交联。然而，光活性鲑鱼降钙素在MCF 7细胞上与一种约80 - 90,000 Mr的成分发生了交联。因此，尽管光活性鲑鱼降钙素能够在所有细胞系中交联类似的受体成分，但辛二酸二琥珀酰亚胺酯的交联能力显然具有细胞特异性。这些数据证实，在迄今为止所检测的细胞系中，降钙素受体包含一种分子量约为85,000 Mr的成分。
