Crystallization and preliminary X-ray diffraction studies of the WW4 domain of the Nedd4-2 ubiquitin-protein ligase.
Ubiquitin-mediated protein modification via covalent attachment of ubiquitin has emerged as one of the most common regulatory processes in all eukaryotes. Nedd4-2, closely related to neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4), is a multimodular ubiquitin-protein ligase comprised of four WW domains and a Hect domain. The WW domains recognize the proline-rich motifs on the multi-subunit amiloride-sensitive epithelial sodium channel (ENaC). To gain insights into the binding of the WW domain to proline-rich peptides, a protein fragment (78 amino acids) containing the fourth WW domain (WW4) of the Nedd4-2 protein was purified and crystallized and X-ray diffraction data were collected. A data set was obtained to 2.5 A resolution from a cryocooled single crystal at a synchrotron source. The crystals belong to the tetragonal space group P4(1)2(1)2 (or P4(3)2(1)2), with unit-cell parameters a = b = 113.43, c = 103.21 A. Analysis of the self-rotation function suggests the presence of four WW4 molecules in the asymmetric unit, with a high unit-cell solvent content of 74%.
Umadevi
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Department of Biochemistry, Case Western Reserve University, Cleveland, OH 44106, USA.
Kumar
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Narayana
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D000584:Amiloride / Q000737:chemistry; D020816:Amino Acid Motifs; D000818:Animals; D018360:Crystallography, X-Ray; D056827:Endosomal Sorting Complexes Required for Transport; D005982:Glutathione Transferase / Q000378:metabolism; D051379:Mice; D015233:Models, Statistical; D000075702:Nedd4 Ubiquitin Protein Ligases; D009474:Neurons / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012997:Solvents; D017356:Synchrotrons; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry*; D014961:X-Ray Diffraction
D056827:Endosomal Sorting Complexes Required for Transport; D010455:Peptides; D012997:Solvents; D011092:Polyethylene Glycols; D000584:Amiloride; D011392:Proline; D000075702:Nedd4 Ubiquitin Protein Ligases; C000618461:Nedd4l protein, mouse; D044767:Ubiquitin-Protein Ligases; D005982:Glutathione Transferase