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Nucleoside-diphosphate kinase from Streptomyces coelicolor
M Brodbeck1, A Rohling, W Wohlleben
1Department of Microbiology, Biozentrum, University of Basel, Switzerland.
European Journal of Biochemistry
|July 1, 1996
Summary
Nucleoside-diphosphate (NDP) kinase from Streptomyces coelicolor was purified and characterized. The enzyme showed unique structural features, including a truncated C-terminus, making it one of the smallest NDP kinases identified.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Nucleoside-diphosphate (NDP) kinases are essential enzymes involved in nucleotide metabolism.
- Streptomyces coelicolor is a well-studied bacterium with a rich source of bioactive compounds.
Purpose of the Study:
- To purify and characterize NDP kinase from Streptomyces coelicolor.
- To investigate the enzyme's kinetic properties and structural features.
- To analyze the gene encoding NDP kinase.
Main Methods:
- Single-step purification using a cAMP binding column.
- Enzyme activity assays and autophosphorylation studies.
- Gel filtration chromatography for molecular mass determination.
- Gene isolation and DNA sequencing.
Main Results:
- NDP kinase was purified to >90% homogeneity with a 3600-fold increase in specific activity.
- The enzyme is a tetramer of a 15-kDa subunit, with an ATP Km of 85 microM.
- Acid lability of the phosphoenzyme intermediate suggests a histidine residue.
- The ndk gene sequence revealed high similarity to other NDP kinases but a unique truncated C-terminus.
Conclusions:
- Streptomyces coelicolor NDP kinase is a small, tetrameric enzyme with distinct structural characteristics.
- The enzyme's properties provide insights into NDP kinase function and evolution.
- The truncated C-terminus represents a novel feature among characterized NDP kinases.