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PMID: 778852 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Cell-free synthesis of rat preproinsulins: characterization and partial amino acid sequence determination.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 73 ·No. 6 ·1976-06-00 ·Pages 1964-8

Chan SJ, Keim P, Steiner DF

Abstract

Whole nucleic acid fractions of isolated rat islets of Langerhans greatly stimulate incorporation of radioactive amino acids into protein in a wheat germ ribosomal system. Approximately 30% of the synthetic product is precipitated with antisera to insulin or proinsulin. Characterization of this material by gel chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicates a molecular mass of 11,500 daltons. Trypsin digestion releases intact A chain as well as tryptic fragments of the C-peptides and B chains of the two rat proinsulins. Automated sequence determination of labeled cell-free product purified by immunoprecipitation discloses the presence of 23 additional amino acids NH2-terminal to the B chain sequence of proinsulin. The partial amino acid sequence of this extension is as follows: NH2-X-Leu (Lys) Met-x-Phe-Leu-Phe-Leu-Leu (Lys) Leu-Leu-x-leu-X-X-X-X-X-X-X-X-proinsulin. On the basis of the above evidence we have designated this peptide preproinsulin.

MeSH Terms

Amino Acid Sequence Animals Cell Nucleus/metabolism Cell-Free System Islets of Langerhans/ultrastructure Molecular Weight Proinsulin/analysis,biosynthesis Protein Biosynthesis RNA, Messenger/metabolism Rats

Chemicals

RNA, Messenger Proinsulin

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Chan S J

Keim P

Steiner D F

References (22)

22 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.73.6.1964

Published

1976-06-00

Pages

1964-8

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC430428

Subset

External Links

PubMed source DOI: 10.1073/pnas.73.6.1964 PMC Full Text View journal details

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Cell-free synthesis of rat preproinsulins: characterization and partial amino acid sequence determination.

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