Crystallization and preliminary X-ray studies of the N-domain of the Wilson disease associated protein.

Abstract

Wilson disease associated protein (ATP7B) is essential for copper transport in human cells. Mutations that affect ATP7B function result in Wilson's disease, a chronic copper toxicosis. Disease-causing mutations within the N-domain of ATP7B (WND) are known to disrupt ATP binding, but a high-resolution X-ray structure of the ATP-binding site has not been reported. The N-domain was modified to delete the disordered loop comprising residues His1115-Asp1138 (WNDDelta(1115-1138)). Unlike the wild-type N-domain, WNDDelta(1115-1138) formed good-quality crystals. Synchrotron diffraction data have been collected from WNDDelta(1115-1138) at the Canadian Light Source. A native WNDDelta(1115-1138) crystal diffracted to 1.7 A resolution and belonged to space group P4(2)2(1)2, with unit-cell parameters a = 39.2, b = 39.2, c = 168.9 A. MAD data were collected to 2.7 A resolution from a SeMet-derivative crystal with unit-cell parameters a = 38.4, b = 38.4, c = 166.7 A. The WNDDelta(1115-1138) structure is likely to be solved by phasing from multiwavelength anomalous diffraction (MAD) experiments.

• Authors: Lili Liu, Christopher O'Grady, Sean A Dalrymple, Lata Prasad, Oleg Y Dmitriev, Louis T J Delbaere
• Journal: Acta crystallographica. Section F, Structural biology and crystallization communications (Acta Crystallogr Sect F Struct Biol Cryst Commun) Vol. 65 Issue Pt 6 Pg. 621-4 (Jun 01 2009) ISSN: 1744-3091 [Electronic] England
• PMID: 19478447 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Cation Transport Proteins
• Copper
• Adenosine Triphosphate
• Adenosine Triphosphatases
• ATP7B protein, human
• Copper-Transporting ATPases

Topics

• Adenosine Triphosphatases (chemistry, genetics, isolation & purification, metabolism)
• Adenosine Triphosphate (metabolism)
• Amino Acid Sequence
• Binding Sites
• Cation Transport Proteins (chemistry, genetics, isolation & purification, metabolism)
• Conserved Sequence
• Copper (chemistry, metabolism)
• Copper-Transporting ATPases
• Crystallization
• Crystallography, X-Ray
• Data Collection
• Escherichia coli (genetics)
• Hepatolenticular Degeneration (genetics)
• Humans
• Models, Molecular
• Molecular Sequence Data
• Nuclear Magnetic Resonance, Biomolecular
• Protein Binding
• Protein Structure, Secondary
• Protein Structure, Tertiary
• Statistics as Topic
• Synchrotrons
• Transformation, Bacterial
• X-Ray Diffraction