De novo biosynthesis of an enzymatically active precursor form of bovine pancreatic RNase

T H Haugen; Edward C Heath

doi:10.1073/pnas.76.6.2689

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De novo biosynthesis of an enzymatically active precursor form of bovine pancreatic RNase

Journal article

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Peer reviewed

De novo biosynthesis of an enzymatically active precursor form of bovine pancreatic RNase

T H Haugen and Edward C Heath

Proceedings of the National Academy of Sciences - PNAS, Vol.76(6), pp.2689-2693

06/1979

DOI: 10.1073/pnas.76.6.2689

PMCID: PMC383673

PMID: 288056

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Abstract

The de novo biosynthesis of RNase (ribonucleate 3'-pyrimidino-oligonucleotidohydrolase, EC 3.1.4.22) was studied in a cell-free rabbit reticulocyte translation system using a polyadenylylated fraction of mRNA isolated from bovine pancreas. Analysis of the [35S]methionine-labeled translation products of pancreas mRNA by polyacrylamide gel electrophoresis indicated the presence of several proteins, one of which corresponded to 16,500 daltons, or approximately 2800 daltons greater than native RNase A. This protein was specifically precipitated from the mixture of products by anti-RNase antibodies. Partial sequence determination of the NH2-terminal region of the anti-RNase antibody-precipitable species indicated that it is a precursor form of RNase A with 25 additional amino acids on its NH2 terminus. The precursor nature of the protein was confirmed by demonstration that a mixture of RNase A and a glycosylated form of the enzyme, RNase B, is formed when translation of the mRNA is conducted in the presence of dog pancreas membranes. Assay of the putative precursor form of RNase for catalytic activity with polycytidylic acid as substrate indicated that the protein has a specific enzymatic activity identical to that of native RNase A.

Glycoproteins - biosynthesis

Antigen-Antibody Complex

Rabbits

Ribonucleases - biosynthesis

Protein Biosynthesis

Animals

Cattle

Glycosides

Kinetics

RNA, Messenger - metabolism

Reticulocytes - metabolism

Pancreas - enzymology

Details

Title: Subtitle: De novo biosynthesis of an enzymatically active precursor form of bovine pancreatic RNase
Creators: T H Haugen
Edward C Heath
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.76(6), pp.2689-2693
DOI: 10.1073/pnas.76.6.2689
PMID: 288056
PMCID: PMC383673
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; United States
Language: English
Date published: 06/1979
Academic Unit: Pathology
Record Identifier: 9984047626602771

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