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Updated: Aug 12, 2026
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Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
Published on: May 10, 2015
Component analysis and characterization of a nuclear deoxyribonucleotidase
1Krebs Institute, Department of Molecular Biology and Biotechnology, University of Sheffield, U.K.
The Biochemical Journal
|March 15, 1994
Summary
Researchers identified a novel
Area of Science:
- Molecular Biology
- Biochemistry
Background:
- Mammalian cells possess a nuclear activity that hydrolyzes deoxyribonucleoside triphosphates (dNTPs).
- Previous studies identified this dNTPase activity and its interaction with dATP, forming a retarded band on gels.
Purpose of the Study:
- To characterize the nature of the retarded species formed during dATP hydrolysis.
- To determine the relationship between the dNTPase activity and the observed complex.
Main Methods:
- Non-denaturing polyacrylamide gel electrophoresis (PAGE).
- Thin-layer chromatography (TLC).
- Gel-filtration chromatography.
- Ion-exchange chromatography.
Main Results:
- The retarded species, termed 'product-nucleotide binding particle' (PNBP*), comprises dATP hydrolysis products bound to an unknown molecule.
- The dNTPase activity is distinct from the PNBP*.
- PNBP* has a molecular mass of 2.5–3 kDa and appears to covalently bind the nucleotide ligand.
- Unliganded PNBP* (or a precursor) is associated with the dNTPase enzyme in the active complex.
Conclusions:
- The study elucidates the composition and properties of the PNBP*, a novel cellular component involved in dNTP metabolism.
- The dNTPase enzyme and PNBP* form a complex that is active in nucleotide hydrolysis.
- Further research is needed to identify the unknown species within PNBP* and its precise biological role.
