Abstract:
:We labeled substance P (SP), neurokinin A (NKA), and [Lys0]gastrin-releasing peptide-27 (GRP) with cyanine 3.18 (cy3). Cy3-peptides purified by HPLC were fully active, determined by [Ca2+]i mobilization. Binding was specific because it was abolished by unlabeled peptides and receptor antagonists. Transfected cells yielded a log-fold greater cy3 intensity than control cells by FACS. Confocal microscopy of transfected cells and cultured enteric neurons showed that cy3-SP bound to surface receptors and was internalized. Internalization was observed in living cells by capture of sequential images. Recovery of surface binding sites was monitored by flow cytometry using cy3-SP. Thus, cy3 neuropeptides can be used to isolate and study receptor-bearing cells.
journal_name
Peptidesjournal_title
Peptidesauthors
Bunnett NW,Dazin PF,Payan DG,Grady EFdoi
10.1016/0196-9781(95)00042-isubject
Has Abstractpub_date
1995-01-01 00:00:00pages
733-40issue
4eissn
0196-9781issn
1873-5169pii
0196-9781(95)00042-Ijournal_volume
16pub_type
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