Importance of the 10-13 region of glucagon for its receptor interactions and activation of adenylate cyclase

John L. Krstenansky, Dev Trivedi, Victor J Hruby

Research output: Contribution to journalArticle

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Abstract

The role of the Tyr10-Ser11-Lys12-Tyr13 region of glucagon in the binding interaction and activation of the glucagon receptor was investigated by means of the synthetic glucagon analogues [Phe13]glucagonamide (2), [Phe10]glucagonamide (3), [Phe10]glucagon (4), [Phe10,13]glucagon (5), [Pro11]glucagon (6), [Pro11,Gly12]glucagonamide (7), [Ala11]glucagon (8), and [Oac11-13]glucagonamide (9). These analogues were synthesized by solid-phase peptide synthesis on p-methylbenzhydrylamine or Merrifield resins with protected Nα-tert-butyloxycarbonyl amino acids. Purification by dialysis, cation-exchange chromatography, gel filtration, and preparative reverse-phase high-performance liquid chromatography (HPLC) gave products that proved homogeneous by thin-layer chromatography and HPLC and on analysis by amino acid analysis, by sequencing, and by α-chymotryptic peptide mapping with HPLC. Biological activities were examined by measurement of the stimulation of liver plasma membrane adenylate cyclase and by specific displacement of [125I]glucagon from glucagon receptors. The results of these studies indicate that while the biological "message" region of glucagon is located elsewhere, the 10-13 region has multiple roles in the glucagon-glucagon receptor interaction: (1) this region provides functional groups for direct binding interaction with the receptor, and (2) this region interacts with the receptor in such a way as to allow the "transduction message" portion of glucagon to interact and activate the receptor.

Original languageEnglish (US)
Pages (from-to)3833-3839
Number of pages7
JournalBiochemistry
Volume25
Issue number13
StatePublished - 1986

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Glucagon Receptors
Glucagon
Adenylyl Cyclases
Chemical activation
High performance liquid chromatography
High Pressure Liquid Chromatography
Amino Acids
Thin layer chromatography
Peptides
Solid-Phase Synthesis Techniques
Peptide Mapping
Dialysis
Protein Sequence Analysis
Reverse-Phase Chromatography
Cell membranes
Thin Layer Chromatography
Chromatography
Bioactivity
Liver
Functional groups

ASJC Scopus subject areas

  • Biochemistry

Cite this

Importance of the 10-13 region of glucagon for its receptor interactions and activation of adenylate cyclase. / Krstenansky, John L.; Trivedi, Dev; Hruby, Victor J.

In: Biochemistry, Vol. 25, No. 13, 1986, p. 3833-3839.

Research output: Contribution to journalArticle

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