Magic angle spinning NMR spectroscopy of thioredoxin reassemblies

作者：Yang Jun; Paramasivan Sivakumar; Marulanda Dabeiba; Cataidi Marcela; Tasayco Maria Luisa; Polenova Tatyana^*

来源：Magnetic Resonance in Chemistry, 2007, 45: S73-S83.

DOI：10.1002/mrc.2092

• solid-state NMR
• resonance assignments
• magic angle spinning
• C-13
• N-15
• thioredoxin
• reassembled thioredoxin
• secondary structure
• tertiary constraints

Differentially isotopically enriched 1-73 (C-13,N-15)/74-108(N-15) and 1-73(N-15)/74-108(C-13,N-15) Escherichia coli thioredoxin reassemblies prepared by fragment complementation were investigated by high-resolution magic angle spinning solid-state NMR spectroscopy. Nearly complete resonance assignments, secondary and tertiary structure analysis are reported for 1-73(C-13,N-15)/74-108(N-15) reassembled thioredoxin. Temperature dependence of the dipolar-assisted rotational resonance (DARR) spectra reveals the residues undergoing intermediate timescale motions at temperatures below -15 degrees C. Analysis of the DARR intensity buildups as a function of mixing time in these reassemblies indicates that at long mixing times medium- and long-range cross-peaks do not experience dipolar truncation, suggesting that isotopic dilution is not required for gaining nontrivial distance restraints for structure calculations.

• 出版日期2007-12