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Yeast tRNAAsp-aspartyl-tRNA synthetase: the crystalline complex
1Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
Journal of Biomolecular Structure & Dynamics
|October 1, 1983
Summary
Yeast aspartyl-tRNA synthetase and its tRNA were crystallized to study their interaction. Neutron diffraction revealed a large interface between the enzyme and tRNA, impacting structural analysis.
Area of Science:
- Biochemistry
- Structural Biology
- Crystallography
Background:
- Aspartyl-tRNA synthetase (AspRS) is crucial for protein synthesis.
- Understanding enzyme-tRNA interactions is key to deciphering translational fidelity.
Purpose of the Study:
- To structurally investigate the yeast AspRS-tRNAAsp complex.
- To determine the stoichiometry and crystal packing of the complex.
Main Methods:
- Co-crystallization of yeast AspRS and tRNAAsp using ammonium sulfate.
- Electrophoresis, biological activity assays, and crystallographic analysis.
- Neutron diffraction with H2O/D2O contrast variation to 20 Å resolution.
Main Results:
- Crystals of AspRS-tRNAAsp complex formed in space group I432 with a 2:1 enzyme-to-tRNA stoichiometry.
- Neutron diffraction data indicated a significant deviation from predicted scattering, suggesting a large, solvent-shielded interface.
- Experimental D2O extinction at 36% differed from the predicted 41%.
Conclusions:
- A substantial interface exists between yeast AspRS and its cognate tRNA.
- This interface region influences solvent accessibility and neutron scattering properties.
- The findings support a model where the enzyme and tRNA interact extensively.