title: Assignment of 1H(N), 15N, 13C(alpha), 13CO and 13C(beta) resonances in a 67 kDa p53 dimer using 4D-TROSY NMR spectroscopy.
abstract: The p53 tumor suppressor is a transcription factor that plays a crucial role in the activation of genes in response to DNA damage. As a first step towards detailed structural studies of the molecule aimed at understanding its regulation, we have used 4D-TROSY triple resonance NMR spectroscopy to obtain nearly complete 1H(N), 15N, 13C(alpha), 13CO and 13C(beta) resonance assignments of a dimeric form of the protein comprising DNA-binding and oligomerization domains (67 kDa). A simple comparison of 4D spectra recorded on p53 molecules consisting of DNA-binding and oligomerization domains with and without the regulatory domain establishes that both constructs have essentially identical chemical shifts. Although the affinity of p53 for target DNA is decreased in constructs containing the regulatory domain, the chemical shift results reported here suggest that this decrease is not due to specific domain interactions involving the regulatory portion of the molecule, or alternatively, that such interactions require the presence of DNA.
authors: Protein Engineering Centers of Excellence and Department of Medical Genetics, The University of Toronto, ON, Canada.
authors: F A
authors: FA
authors: Mulder
authors: A
authors: A
authors: Ayed
authors: D
authors: D
authors: Yang
authors: C H
authors: CH
authors: Arrowsmith
authors: L E
authors: LE
authors: Kay
mesh_terms: D001665:Binding Sites; D002247:Carbon Isotopes; D002248:Carbon Monoxide; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D019281:Dimerization; D006859:Hydrogen; D046911:Macromolecular Substances; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D046911:Macromolecular Substances; D009587:Nitrogen Isotopes; D016159:Tumor Suppressor Protein p53; D002248:Carbon Monoxide; D006859:Hydrogen; D004247:DNA