{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T10:59:35Z","timestamp":1760266775103},"reference-count":21,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2009,1,29]],"date-time":"2009-01-29T00:00:00Z","timestamp":1233187200000},"content-version":"vor","delay-in-days":578,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"published-print":{"date-parts":[[2007,7]]},"abstract":"<jats:sec><jats:title>Background and purpose:<\/jats:title><jats:p>Transmembrane proline (P) residues in family A G protein\u2010coupled receptors (GPCRs) form functionally important kinks in their helices. These residues are little studied in family B GPCRs but experiments with the VPAC1 receptor and calcitonin receptor\u2010like receptor (CL) show parallels with family A receptors. We sought to determine the function of these residues in the insert negative form of the human calcitonin receptor, a close relative of CL.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental approach:<\/jats:title><jats:p>Proline residues within the transmembrane domains of the calcitonin receptor (P246, P249, P280, P326, P336) were individually mutated to alanine (A) using site\u2010directed mutagenesis. Receptors were transiently transfected into Cos\u20107 cells using polyethylenimine and salmon and human calcitonin\u2010induced cAMP responses measured. Salmon and human calcitonin competition binding experiments were also performed and receptor cell\u2010surface expression assessed by whole cell ELISA.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key results:<\/jats:title><jats:p>P246A, P249A and P280A were wild\u2010type in terms of human calcitonin\u2010induced cAMP activation. P326A and P336A had reduced function (165 and 12\u2010fold, respectively). In membranes, human calcitonin binding was not detectable for any mutant receptor but in whole cells, binding was detected for all mutants apart from P326A. Salmon calcitonin activated mutant and wild\u2010type receptors equally, although B<jats:sub>max<\/jats:sub> values were reduced for all mutants apart from P326A.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Implications:<\/jats:title><jats:p>P326 and P336 are important for the function of human calcitonin receptors and are likely to be involved in generating receptor conformations appropriate for agonist binding and receptor activation. However, agonist\u2010specific effects were observed, implying distinct conformations of the human calcitonin receptor.<\/jats:p><jats:p><jats:italic>British Journal of Pharmacology<\/jats:italic> (2007) <jats:bold>151<\/jats:bold>, 678\u2013687; doi:<jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" ext-link-type=\"doi\" xlink:href=\"10.1038\/sj.bjp.0707246\">10.1038\/sj.bjp.0707246<\/jats:ext-link><\/jats:p><\/jats:sec>","DOI":"10.1038\/sj.bjp.0707246","type":"journal-article","created":{"date-parts":[[2007,5,8]],"date-time":"2007-05-08T10:54:06Z","timestamp":1178621646000},"page":"678-687","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Agonist\u2010dependent consequences of proline to alanine substitution in the transmembrane helices of the calcitonin receptor"],"prefix":"10.1111","volume":"151","author":[{"given":"R 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