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Endocrinology, Vol 113, 711-719, Copyright © 1983 by Endocrine Society

ARTICLES

Monoiodoglucagon: synthesis, purification by high pressure liquid chromatography, and characteristics as a receptor probe

FJ Rojas, TL Swartz, R Iyengar, AJ Garber and L Birnbaumer

The synthesis of [125I-Tyr10]monoiodoglucagon from glucagon and carrier- free 125I using 1,3,4,6-tetrachloro-3-6-diphenylglycouril (Iodogen) and its separation in pure form by reverse phase high pressure liquid chromatography (HPLC) over C18-muBondapak columns using two consecutive linear gradients between solvent A [40:60 mixture of methanol and 10 mM H3PO4 in H2O (pH adjusted to 3.0 with triethylamine)] and solvent B (50:50 mixture of acetonitrile and 0.1 M Tris-HCl, pH 9.0) is reported. The newly synthesized [125I]monoiodoglucagon is shown to activate adenylyl cyclase in liver membranes with an EC50 between 5- and 8-fold lower than that of native glucagon. Further, it binds specifically to sites on liver plasma membranes that have the characteristics of glucagon receptors in terms of guanine nucleotide sensitivity and rates of reaction. It is suggested that [125I-Tyr10]monoiodoglucagon is a suitable probe for studying structural and functional properties of glucagon receptors.


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A Yatani, J Codina, Y Imoto, J. Reeves, L Birnbaumer, and A. Brown
A G protein directly regulates mammalian cardiac calcium channels
Science, November 27, 1987; 238(4831): 1288 - 1292.
[Abstract] [PDF]


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