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Biochemical characterization of mouse brain necdin
1Department of Molecular Neurobiology, Tokyo Metropolitan Institute for Neuroscience, Japan.
The Biochemical Journal
|March 15, 1996
Summary
Necdin is a nuclear basic DNA-binding protein identified in mouse brain. It forms large complexes and associates with molecules to regulate transcriptionally active genes and nuclear function.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Necdin is a protein encoded by neural differentiation-specific mRNA.
- It is derived from embryonal carcinoma cells (P19).
Purpose of the Study:
- To characterize the mouse brain necdin protein.
- To investigate its DNA-binding properties and interactions within the nucleus.
Main Methods:
- Western blotting and silver-staining analysis using affinity-purified antibodies.
- Gel filtration chromatography (Sephacryl S-300) and sucrose gradient centrifugation.
- DNA-affinity chromatography and isoelectric focusing.
Main Results:
- Necdin has a molecular mass of 51 kDa and localizes to nucleosomal fractions.
- It forms large complexes (peak at 700 kDa) and binds tightly to double-stranded DNA.
- Purified necdin (pI 9.1) is a basic DNA-binding protein involved in nucleosomal complexes.
Conclusions:
- Necdin is a nuclear basic DNA-binding protein.
- It associates with other molecules to form complexes that regulate transcriptionally active genes.
- Necdin plays a role in nuclear function and gene regulation.