Biotinyl analogues of amylin as biologically active probes for amylin/CGRP receptor recognition.
Biotinyl analogues of rat amylin were synthesised with sulfosuccinimidyl 2-(biotinamido) ethyl-1,3-dithiopropionate (NHS-SS-Biotin). Biotinylated amylin peptides were purified by HPLC, quantitated, and the presence of the biotin group at Lys-1 confirmed by peroxidase-labelled avidin and FAB mass spectroscopy. Amylin-biotin retained a similar affinity for binding to rat liver plasma membranes compared with rat amylin and also completely inhibited insulin-stimulated glycogen synthesis in rat soleus muscle incubated in vitro. These biologically active amylin probes will enable a complete analysis of amylin/CGRP receptor expression in various cell types and facilitate the isolation and characterisation of the hormone-receptor complex.
Chantry
A
A
Department of Biochemistry, University of Oxford, UK.
Foot
E A
EA
Leighton
B
B
Day
A J
AJ
Willis
A C
AC
D000682:Amyloid / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001710:Biotin / Q000737:chemistry; D002116:Calcitonin / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D006003:Glycogen / Q000096:biosynthesis; D007124:Immunoenzyme Techniques; D007328:Insulin / Q000494:pharmacology; D058228:Islet Amyloid Polypeptide; D008099:Liver / Q000166:cytology; D015335:Molecular Probes / Q000378:metabolism; D009132:Muscles / Q000187:drug effects / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D018003:Receptors, Calcitonin; D011956:Receptors, Cell Surface / Q000378:metabolism*; D016339:Spectrometry, Mass, Fast Atom Bombardment
D000682:Amyloid; D007328:Insulin; D058228:Islet Amyloid Polypeptide; D015335:Molecular Probes; D010446:Peptide Fragments; D018003:Receptors, Calcitonin; D011956:Receptors, Cell Surface; D001710:Biotin; D006003:Glycogen; D002116:Calcitonin