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.
Original language | English (US) |
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Pages (from-to) | 733-740 |
Number of pages | 8 |
Journal | Peptides |
Volume | 16 |
Issue number | 4 |
DOIs | |
State | Published - 1995 |
Keywords
- Endocytosis
- Flow analysis
- Fluorescent peptides
- Gastrin-releasing peptide
- Neurokinin A
- Substance P
ASJC Scopus subject areas
- Biochemistry
- Physiology
- Endocrinology
- Cellular and Molecular Neuroscience