Abstract
Ube3A (also referred to as E6AP for E6 Associated Protein) is a E3 ubiquitin-protein ligase implicated in the development of Angelman syndrome by controlling degradation of synaptic protein Arc and oncogenic papilloma virus infection by controlling degradation of p53. This article describe the solution NMR structure of the conserved N-terminal domain of human Ube3A (residues 24-87) that contains two residues (Cys44 and Arg62) found to be mutated in patients with Angelman syndrome. The structure of this domain adopts a novel Zn-binding fold we called AZUL (Amino-terminal Zn-finger of Ube3a Ligase). The AZUL domain has a helix-loop-helix architecture with a Zn ion coordinated by four Cys residues arranged in Cys-X4-Cys-X4-Cys-X28-Cys motif. Three of the Zn-bound residues are located in a 23-residue long and well structured loop that connects two α-helicies.
Similar content being viewed by others
References
Alberts IL, Nadassy K, Wodak SJ (1998) Analysis of zinc binding sites in protein crystal structures. Protein Sci 7(8):1700–1716
Bai JL, Qu YJ et al (2011) A novel missense mutation of the ubiquitin protein ligase E3A gene in a patient with Angelman syndrome. Chin Med J Engl 124(1):84–88
Beaudenon S, Huibregtse JM (2008) HPV E6, E6AP and cervical cancer. BMC Biochem 9 suppl 1:S4.
Bhattacharya A, Tejero R, Montelione GT (2007) Evaluating protein structures determined by structural genomics consortia. Proteins 66(4):778–795
Brünger AT, Adams PD et al (1998) Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr D Biol Crystallogr 54(Pt 5):905–921
Cooper EM, Hudson AW et al (2004) Biochemical analysis of Angelman syndrome-associated mutations in the E3 ubiquitin ligase E6-associated protein. J Biol Chem 279(39):41208–41217
Delaglio F, Grzesiek S et al (1995) Nmrpipe—a multidimensional spectral processing system based on unix Pipes. J Biomol NMR 6(3):277–293
Goddard TD, Kneller DG (2003) Sparky—NMR assignment and integration software.
Greer PL, Hanayama R et al (2010) The Angelman syndrome protein Ube3A regulates synapse development by ubiquitinating arc. Cell 140(5):704–716
Güntert P (2004) Automated NMR structure calculation with CYANA. Methods Mol Biol 278:353–378
Holm L, Rosenström P (2010) Dali server: conservation mapping in 3D. Nucleic Acids Res 38(Web Server issue): W545–W549
Huang YJ, Powers R, Montelione GT (2005) Protein NMR recall, precision, and F-measure scores (RPF scores): structure quality assessment measures based on information retrieval statistics. J Am Chem Soc 127:1665–1674
Kay LE (1995) Pulsed field gradient multi-dimensional NMR methods for the study of protein structure and dynamics in solution. Prog Biophys Mol Biol 63(3):277–299
Koradi R, Billeter M and Wuthrich K (1996) MOLMOL: a program for display and analysis of macromolecular structures. J Mol Graph 14(1):51–55, 29–32.
Kornhaber GJ, Snyder D, Moseley HNB, Montelione GT (2006) Identification of zinc-ligated cysteine residues based on C-13 alpha and C-13 beta chemical shift data. J Biomol NMR 34(4):259–269
Lemak A, Gutmanas A et al (2011) A novel strategy for NMR resonance assignment and protein structure determination. J Biomol NMR 49(1):27–38
Linge JP, Williams MA et al (2003) Refinement of protein structures in explicit solvent. Proteins 50(3):496–506
Malzac P, Webber H et al (1998) Mutation analysis of UBE3A in Angelman syndrome patients. Am J Hum Genet 62(6):1353–1360
Shen Y, Lange O et al (2008) Consistent blind protein structure generation from NMR chemical shift data. Proc Natl Acad Sci USA 105(12):4685–4690
Walboomers JM, Jacobs MV et al (1999) Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 189(1):12–19
Williams CA, Driscoll DJ, Dagli AI (2010) Clinical and genetic aspects of Angelman syndrome. Genet Med 12(7):385–395
Yee A, Chang X et al (2002) An NMR approach to structural proteomics. Proc Natl Acad Sci USA 99(4):1825–1830
Acknowledgments
Authors are thankful to Peter Loppnau for Ube3A constructs cloning. This work is supported by the US National Institute of Health Protein Structure Initiative (P50-GM62413-01 and GM67965) through the Northeast Structural and the Structural Genomics Consortium a registered charity (number 1097737) that receives funds from the Canadian Institutes of Health Research, the Canadian Foundation for Innovation, Eli Lilly, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Novartis, the Ontario Minstry of Research and Innovation, Pfizer, and the Wellcome Trust. CHA holds a Canada Research Chair in Structural Proteomics. IB is supported by grant 1P30GM092369 from the NIH.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Lemak, A., Yee, A., Bezsonova, I. et al. Zn-binding AZUL domain of human ubiquitin protein ligase Ube3A. J Biomol NMR 51, 185 (2011). https://doi.org/10.1007/s10858-011-9552-y
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10858-011-9552-y