Identification of the active amino acid residue of the polypeptide of ATP-dependent protein breakdown

The heat-stable polypeptide of the ATP-dependent proteolytic system was previously found to form covalent conjugates with proteins and to be activated by ATP in an adenylylation mechanism. To identify the functional amino acid of the polypeptide, the activated residue was specifically labeled by the...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1981-02, Vol.256 (4), p.1525-1528
Main Authors: Hershko, A, Ciechanover, A, Rose, I A
Format: Article
Language:English
Subjects:
Adenosine Triphosphate
Amino Acids - analysis
Animals
Borohydrides
Chromosomal Proteins, Non-Histone - blood
Erythrocytes - analysis
Glycine - analysis
Humans
Rabbits
Reticulocytes - enzymology
Tritium
Ubiquitins
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The heat-stable polypeptide of the ATP-dependent proteolytic system was previously found to form covalent conjugates with proteins and to be activated by ATP in an adenylylation mechanism. To identify the functional amino acid of the polypeptide, the activated residue was specifically labeled by the reductive cleavage of the intermediate with [3H]borohydride. Following acid hydrolysis, the reduced labeled derivative was found to be completely oxidizable by periodate with formation of [3H]formaldehyde, and was identified as ethanolamine by thin layer chromatography, electrophoresis, and amino acid analyzer chromatography. These results indicate that the activated amino acid residue of the polypeptide is COOH-terminal glycine.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)69833-9