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A new strategy for sequential assignment of intrinsically unstructured proteins based on 15N single isotope labelling.
Lopez, Juan; Ahuja, Puneet; Gerard, Melanie; Wieruszeski, Jean-Michel; Lippens, Guy.
Afiliación
  • Lopez J; CNRS UMR 8576, Unité de Glycobiologie Structurale et Fonctionnelle, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France.
J Magn Reson ; 236: 1-6, 2013 Nov.
Article en En | MEDLINE | ID: mdl-24018100
We describe a new efficient strategy for the sequential assignment of amide resonances of a conventional (15)N-(1)H HSQC spectrum of intrinsically unfolded proteins, based on composite NOESY-TOCSY and TOCSY-NOESY mixing times. These composite mixing times lead to a Hα-proton mediated unidirectional transfer of amide to amide proton. We have implemented the composite mixing times in an HSQC-NOESY-HSQC manner to obtain directional connectivity between amides of neighbouring residues. We experimentally determine the optimal mixing times for both transfer schemes, and demonstrate its use in the assignment for both a fragment of the neuronal tau protein and for α-synuclein.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas / Marcaje Isotópico Idioma: En Revista: J Magn Reson Asunto de la revista: DIAGNOSTICO POR IMAGEM Año: 2013 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas / Marcaje Isotópico Idioma: En Revista: J Magn Reson Asunto de la revista: DIAGNOSTICO POR IMAGEM Año: 2013 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Estados Unidos