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Mouse testis-brain RNA-binding protein (TB-RBP): expression, purification and crystal X-ray diffraction
J M Pascal1, V M Chennathukuzhi, N B Hecht
1Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, TX 78712, USA. jrobertus@mail.utexas.edu
Acta Crystallographica. Section D, Biological Crystallography
|October 27, 2001
Summary
Testis-brain RNA-binding protein (TB-RBP) regulates mRNA expression in the brain and testis. This study details TB-RBP expression, purification, crystallization, and its role in RNA translocation and cell development.
Area of Science:
- Molecular Biology
- Cell Biology
- Structural Biology
Background:
- Testis-brain RNA-binding protein (TB-RBP) is crucial for temporal and spatial mRNA expression control.
- TB-RBP is abundant in the brain and male germ cells, regulating mRNA translation during development.
- TB-RBP-mRNA complexes interact with microtubules, facilitating RNA translocation.
Purpose of the Study:
- To describe the expression, purification, and crystallization of TB-RBP.
- To present preliminary X-ray diffraction data for TB-RBP.
- To investigate the multimeric state of TB-RBP using dynamic light scattering.
Main Methods:
- Expression and purification of TB-RBP.
- Crystallization of TB-RBP/translin complex.
- X-ray diffraction analysis.
- Dynamic light scattering (DLS) for multimeric state determination.
Main Results:
- TB-RBP was successfully expressed, purified, and crystallized in space group P2(1)2(1)2.
- Preliminary X-ray diffraction data were obtained.
- Dynamic light scattering provided insights into the multimeric state of TB-RBP.
Conclusions:
- The study provides foundational structural and biophysical data for TB-RBP.
- Understanding TB-RBP structure and function is key to its role in mRNA regulation.
- Further structural studies will elucidate TB-RBP's mechanism in RNA translocation and development.