A new strategy for sequential assignment of intrinsically unstructured proteins based on N-15 single isotope labelling

作者：Lopez Juan; Ahuja Puneet; Gerard Melanie; Wieruszeski Jean Michel; Lippens Guy^*

来源：Journal of Magnetic Resonance, 2013, 236: 1-6.

DOI：10.1016/j.jmr.2013.07.007

• Assignment
• Intrinsically unstructured protein
• NOESY
• TOCSY

We describe a new efficient strategy for the sequential assignment of amide resonances of a conventional N-15-H-1 HSQC spectrum of intrinsically unfolded proteins, based on composite NOESY-TOCSY and TOCSY-NOESY mixing times. These composite mixing times lead to a Hoc-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 alpha-synuclein.

• 出版日期2013-11
• 单位KU Leuven