In vitro biosynthesis and processing of immunologically identified methionine-enkephalin precursor protein

S. L. Sabol, C. M. Liang, Satya Dandekar, L. S. Kranzler

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The biosynthesis and initial processing of the methionine-enkephalin precursor preproenkephalin A were examined by cell-free translation of mRNA from brain and adrenal medulla. A novel antiserum raised against Met-enkephalin-Arg6-Phe7 was shown to react with bovine adrenal medulla fractions (apparent M(r) 34,000) containing proenkephalin A. Affinity-purified antibodies from this antiserum were used to immunoprecipitate cell-free translated [35S]Met-enkephalin-containing protein. A protein of apparent M(r) 30,000 ± 500 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was the only Met-enkephalin precursor consistently synthesized by translation of mRNA from bovine or guinea pig striatum, rat brain, or bovine adrenal medulla. The presence of [35S]Met-enkephalin sequences in this protein was confirmed by high pressure liquid chromatography of trypsin/carboxypeptidase B digests. Dog pancreas endoplasmic reticulum membranes converted the M(r) 30,000 protein to an immunoreactive protein of apparent M(r) 28,500 that lacked significant co re glycosylation. These results suggest that a protein similar or identical to bovine adrenal medullary preproenkephalin A is the major Met-enkephalin precursor synthesized in brain as well as adrenal medulla, and preproenkephalin A is converted to a protein resembling proenkephalin A, presumably by removal of a signal peptide.

Original languageEnglish (US)
Pages (from-to)2697-2704
Number of pages8
JournalJournal of Biological Chemistry
Volume258
Issue number4
StatePublished - 1983
Externally publishedYes

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Methionine Enkephalin
Protein Precursors
Biosynthesis
Adrenal Medulla
Processing
Proteins
Brain
Protein Biosynthesis
Immune Sera
Carboxypeptidase B
High pressure liquid chromatography
Glycosylation
Messenger RNA
Antibody Affinity
Protein Sorting Signals
In Vitro Techniques
Electrophoresis
Sodium Dodecyl Sulfate
Endoplasmic Reticulum
Trypsin

ASJC Scopus subject areas

  • Biochemistry

Cite this

In vitro biosynthesis and processing of immunologically identified methionine-enkephalin precursor protein. / Sabol, S. L.; Liang, C. M.; Dandekar, Satya; Kranzler, L. S.

In: Journal of Biological Chemistry, Vol. 258, No. 4, 1983, p. 2697-2704.

Research output: Contribution to journalArticle

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