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2ojr

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{{STRUCTURE_2ojr| PDB=2ojr | SCENE= }}
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'''Structure of ubiquitin solved by SAD using the Lanthanide-Binding Tag'''
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===Structure of ubiquitin solved by SAD using the Lanthanide-Binding Tag===
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==Overview==
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A double-lanthanide-binding tag (dLBT), a small peptide sequence engineered to bind two lanthanide ions (e.g., Tb3+) with high affinity, was used to solve the phase problem for the structure determination of ubiquitin by the single-wavelength anomalous diffraction (SAD) method. Since the dLBT is comprised exclusively of encoded amino acids, the necessity for the incorporation of unnatural amino acids or chemical modification of the protein as a prerequisite for X-ray structure determination is eliminated. A construct encoding the dLBT as an N-terminal fusion with ubiquitin provides for facile expression and purification using standard methods. Phasing of the single-wavelength X-ray data (at 2.6 A resolution) using only the anomalous signal from the two tightly bound Tb3+ ions in the dLBT led to clear electron-density maps. Nearly 75% of the ubiquitin structure was built using automated model-building software without user intervention. It is anticipated that this technique will be broadly applicable, complementing existing macromolecular phasing methodologies. The dLBT should be particularly useful in cases where protein derivatization with heavy atoms proves to be problematic or synchrotron facilities are unavailable.
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{{ABSTRACT_PUBMED_17497863}}
==About this Structure==
==About this Structure==
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 09:51:37 2008''

Revision as of 06:51, 28 July 2008

Image:2ojr.png

Template:STRUCTURE 2ojr

Structure of ubiquitin solved by SAD using the Lanthanide-Binding Tag

Template:ABSTRACT PUBMED 17497863

About this Structure

2OJR is a Single protein structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.

Reference

Double-lanthanide-binding tags for macromolecular crystallographic structure determination., Silvaggi NR, Martin LJ, Schwalbe H, Imperiali B, Allen KN, J Am Chem Soc. 2007 Jun 6;129(22):7114-20. Epub 2007 May 12. PMID:17497863

Page seeded by OCA on Mon Jul 28 09:51:37 2008

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