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Isolation and characterization of a cDNA clone encoding rat nucleoside diphosphate kinase
N Kimura1, N Shimada, K Nomura
1Department of Biochemistry, Tokyo Metropolitan Institute of Gerontology, Japan.
The Journal of Biological Chemistry
|September 15, 1990
Summary
Researchers isolated a cytosolic nucleoside diphosphate (NDP) kinase cDNA from rat muscle. The mature enzyme has 147 amino acids and is widely distributed across tissues.
Area of Science:
- Molecular Biology
- Biochemistry
- Enzymology
Background:
- Cytosolic nucleoside diphosphate (NDP) kinase is an essential enzyme involved in nucleotide metabolism.
- Understanding the structure and distribution of NDP kinase is crucial for comprehending its biological roles.
Purpose of the Study:
- To isolate and characterize the cDNA clone for rat skeletal muscle cytosolic NDP kinase.
- To determine the primary amino acid sequence and molecular weight of the enzyme.
- To investigate the tissue-specific expression of NDP kinase mRNA.
Main Methods:
- cDNA library screening using synthetic oligonucleotide probes.
- Nucleotide sequencing to determine the cDNA sequence.
- Amino acid sequence analysis to deduce protein structure.
- Northern blot hybridization to analyze mRNA distribution.
Main Results:
- A 621-base pair cDNA clone encoding cytosolic NDP kinase was isolated.
- The deduced protein sequence indicated a mature enzyme of 147 amino acids (16,724 Da) after post-translational modification.
- NDP kinase mRNA was detected in various rat tissues, with consistent mRNA size across all tissues examined.
Conclusions:
- The study provides the complete primary structure of rat cytosolic NDP kinase.
- The widespread tissue distribution of NDP kinase mRNA suggests a ubiquitous role for the enzyme.
- The findings contribute to understanding the molecular basis of nucleotide metabolism and enzyme function.