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Crystals of acetylated SecB diffract to 2.3-A resolution
C Dekker1, B de Kruijff, G de Korte-Kool
1Department of Biochemistry of Membranes, Department of Crystal and Structural Chemistry, Institute of Biomembranes, Bijvoet Center for Biomolecular Research, Padualaan 8, Utrecht, NL-3584 CH, The Netherlands.
Journal of Structural Biology
|January 14, 2000
Summary
The molecular chaperone SecB, crucial for protein translocation in Escherichia coli, was crystallized in an acetylated form. This acetylation may explain the high-resolution diffraction achieved in this study.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- SecB is a molecular chaperone essential for protein translocation in Escherichia coli.
- Understanding SecB's structure is key to elucidating its function in protein transport.
Purpose of the Study:
- To purify and crystallize the molecular chaperone SecB from an overproducing strain.
- To determine the structural characteristics of crystallized SecB, including post-translational modifications.
Main Methods:
- Overexpression and purification of SecB from Escherichia coli.
- Crystallization of purified SecB.
- X-ray diffraction analysis of SecB crystals.
- Electrospray ionization mass spectrometry (ESI-MS) for structural analysis.
Main Results:
- SecB was successfully purified and crystallized, yielding crystals that diffracted to 2.3-A resolution.
- Mass spectrometry analysis revealed that the crystallized SecB is an acetylated form.
- Sequence analysis indicated N-terminal acetylation in vivo, suggesting deacetylation may occur during purification.
Conclusions:
- The observed N-terminal acetylation of SecB may contribute to the high resolution of the obtained crystals.
- This finding provides insights into SecB's structural integrity and potential modifications affecting its properties.