Carboxyfluorescein and biotin neuromedin C analogues: synthesis and applications.
Two neuromedin C (NC) analogues were constructed by Fmoc synthesis and in situ coupling of 4(5)-carboxyfluorescein or biotin to the N-terminus. Both displayed full agonism in an amylase release assay and cross-reacted fully with a NC-specific antiserum. Biotin NC functioned in a streptavidin-capture ELISA. Carboxyfluorescein NC was used to probe receptor localization in rat stomach. Specific NC binding sites, which did not interact with substance P, angiotensin I, or neurokinin A, were labeled in the antrum. Identity of NC binding sites was confirmed by microautoradiography. The specifically labeled cells were all found in the lamina propria and at least some of cells were identified as eosinophils.
Walker
B
B
Division of Biochemistry, Queen's University of Belfast, N. Ireland, UK.
Gray
J
J
Burns
D M
DM
Wang
Q
Q
Adrian
T E
TE
Nichols
D H
DH
Murphy
R F
RF
Nelson
J
J
D000681:Amylases / Q000378:metabolism*; D000818:Animals; D000918:Antibody Specificity; D001667:Binding, Competitive; D001710:Biotin; D001839:Bombesin / Q000031:analogs & derivatives* / Q000032:analysis / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D005452:Fluoresceins; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007668:Kidney; D010179:Pancreas / Q000187:drug effects / Q000201:enzymology*; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism / Q000494:pharmacology*; D018004:Receptors, Bombesin / Q000032:analysis / Q000096:biosynthesis; D014162:Transfection
D005452:Fluoresceins; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D018004:Receptors, Bombesin; C024098:6-carboxyfluorescein; D001710:Biotin; C040794:neuromedin C; D000681:Amylases; D001839:Bombesin