Glucagon precursors identified by immunoprecipitation of products of cell-free translation of messenger RNA

P. K. Lund, Richard Goodman, J. W. Jacobs, J. F. Habener

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Polyadenylated RNA extracted from anglerfish islets was translated in a wheat germ cell-free system containing [35S]methionine in the presence and absence of microsomal membranes prepared from a canine pancreas. Labeled translation products were analyzed by immunoprecipitation with an antiserum to porcine glucagon, followed by electrophoresis of the translation products and immunoprecipitated proteins on SDS polyacrylamide gels. In the absence of microsomal membranes two proteins of M(r) = 14.500 and M(r) = 12,500 were specifically immunoprecipitated with antiglucagon serum. Addition of microsomal membranes to the translation reactions resulted in a diminution of the labeled protein of M(r) = 14,500 and a marked increase in the immunoreactive protein of M(r) = 12,500. The protein of M(r) = 12,500 was resistant to degradation by proteolytic enzymes added to translation reactions, indicating that it was segregated within microsomal vesicles. These results are consistent with synthesis of anglerfish islet glucagon in the form of a pre-prohormonal precursor (M(r) = 14,500) containing a leader sequence that is cotranslationally cleaved from the protein by enzymes associated with microsomal membranes to produce a smaller intermediate prohormonal precursor (M(r) = 12,500) of pancreatic glucagon (M(r) = 3500).

Original languageEnglish (US)
Pages (from-to)583-586
Number of pages4
JournalDiabetes
Volume29
Issue number7
StatePublished - 1980
Externally publishedYes

Fingerprint

Proglucagon
Immunoprecipitation
Messenger RNA
Glucagon
Proteins
Membranes
Cell-Free System
Germ Cells
Methionine
Triticum
Electrophoresis
Canidae
Immune Sera
Pancreas
Membrane Proteins
Peptide Hydrolases
Swine
Enzymes
Serum

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Internal Medicine

Cite this

Glucagon precursors identified by immunoprecipitation of products of cell-free translation of messenger RNA. / Lund, P. K.; Goodman, Richard; Jacobs, J. W.; Habener, J. F.

In: Diabetes, Vol. 29, No. 7, 1980, p. 583-586.

Research output: Contribution to journalArticle

Lund, P. K. ; Goodman, Richard ; Jacobs, J. W. ; Habener, J. F. / Glucagon precursors identified by immunoprecipitation of products of cell-free translation of messenger RNA. In: Diabetes. 1980 ; Vol. 29, No. 7. pp. 583-586.
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