Characterization of receptors using cyanine 3-labeled neuropeptides.

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

Peptides

journal_title

Peptides

authors

Bunnett NW,Dazin PF,Payan DG,Grady EF

doi

10.1016/0196-9781(95)00042-i

subject

Has Abstract

pub_date

1995-01-01 00:00:00

pages

733-40

issue

4

eissn

0196-9781

issn

1873-5169

pii

0196-9781(95)00042-I

journal_volume

16

pub_type

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