| Products of the inactivation of ribonucleoside diphosphate reductase from Escherichia coli with 2'-azido-2'-deoxyuridine 5'-diphosphate. | |
| | |
MedLine Citation:
|
PMID: 3307907 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Ribonucleoside diphosphate reductase (RDPR) from Escherichia coli was completely inactivated by 1 equiv of the mechanism-based inhibitor 2'-azido-2'-deoxyuridine 5'-diphosphate (N3UDP). Incubation of RDPR with [3'-3H]N3UDP resulted in 0.2 mol of 3H released to solvent per mole of enzyme inactivated, indicating that cleavage of the 3' carbon-hydrogen bond occurred in the reaction. Incubation of RDPR with [beta-32P]N3UDP resulted in stoichiometric production of inorganic pyrophosphate. One equivalent of uracil was eliminated from N3UDP, but no azide release was detected. Analysis of the reaction of RDPR with [15N3]N3UDP by mass spectrometry revealed that the azide moiety was converted to 0.9 mol of nitrogen gas per mole of enzyme inactivated. The tyrosyl radical of the B2 subunit was destroyed during the inactivation by N3UDP as reported previously [Sjöberg, B.-M., Gräslund, A., & Eckstein, F. (1983) J. Biol. Chem. 258, 8060-8067], while the specific activity of the B1 subunit was reduced by half. Incubation of [5'-3H]N3UDP with RDPR resulted in stoichiometric covalent radiolabeling of the enzyme. Separation of the enzyme's subunits by chromatofocusing revealed that the modification was specific for the B1 subunit. |
| | |
Authors:
|
S P Salowe; M A Ator; J Stubbe |
Related Documents
:
|
12604197 - Ligand-induced conformational changes of betaine aldehyde dehydrogenase from pseudomona... 1476707 - Kinetics of inactivation of 4-aminobutyrate aminotransferase by 3-bromopyruvate. 6247337 - Fluoride, pyrophosphate, and base release from 2'-deoxy-2'-fluoronucleoside 5'-diphosph... 8448127 - Role of cysteines in the activation and inactivation of brewers' yeast pyruvate decarbo... 16540087 - Isolation of a ribonuclease from sanchi ginseng (panax pseudoginseng) flowers distinct ... 20386667 - Covalent anchoring of chloroperoxidase and glucose oxidase on the mesoporous molecular ... |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
|
Title: Biochemistry Volume: 26 ISSN: 0006-2960 ISO Abbreviation: Biochemistry Publication Date: 1987 Jun |
Date Detail:
|
Created Date: 1987-11-16 Completed Date: 1987-11-16 Revised Date: 2007-11-14 |
Medline Journal Info:
|
Nlm Unique ID: 0370623 Medline TA: Biochemistry Country: UNITED STATES |
Other Details:
|
Languages: eng Pagination: 3408-16 Citation Subset: IM |
Affiliation:
|
Department of Biochemistry, College of Agriculture and Life Sciences, University of Wisconsin-Madison 53706. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Affinity Labels
/
pharmacology* Azides / pharmacology* Deoxyuracil Nucleotides / pharmacology* Escherichia coli / enzymology* Hydroxyurea / pharmacology Kinetics Protein Binding Ribonucleoside Diphosphate Reductase / antagonists & inhibitors* Ribonucleotide Reductases / antagonists & inhibitors* Tritium |
| Grant Support | |
ID/Acronym/Agency:
|
AM 01222/AM/NIADDK NIH HHS; GM 29595/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Affinity Labels; 0/Azides; 0/Deoxyuracil Nucleotides; 10028-17-8/Tritium; 127-07-1/Hydroxyurea; 36792-49-1/2'-azido-2'-deoxyuridine 5'-diphosphate; EC 1.17.4.-/Ribonucleotide Reductases; EC 1.17.4.1/Ribonucleoside Diphosphate Reductase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Kinetic mechanism of histidinol dehydrogenase: histidinol binding and exchange reactions.
Next Document: Pyridine coenzyme analogues. Synthesis and characterization of alpha- and beta-nicotinamide arabinos...