Band-selective 13C homonuclear 3D spectroscopy for solid proteins at high field with rotor-synchronized soft pulses

Donghua H. Zhou, Kathryn D. Kloepper, Kem A. Winter, Chad M. Rienstra

Research output: Contribution to journalArticlepeer-review

Abstract

We demonstrate improved 3D 13C-13C-13C chemical shift correlation experiments for solid proteins, utilizing band-selective coherence transfer, scalar decoupling and homonuclear zero-quantum polarization transfer. Judicious use of selective pulses and a z-filter period suppress artifacts with a two-step phase cycle, allowing higher digital resolution in a fixed measurement time. The novel correlation of Cali-Cali-CX (Cali for aliphatic carbons, CX for any carbon) reduces measurement time by an order of magnitude without sacrificing digital resolution. The experiment retains intensity from side-chain carbon resonances whose chemical shift dispersion is critical to minimize spectral degeneracy for large proteins with a predominance of secondary structure, such β-sheet rich fibrillar proteins and α-helical membrane proteins. We demonstrate the experiment for the β1 immunoglobulin binding domain of protein G (GB1) and fibrils of the A30P mutant of α-synuclein, which is implicated in Parkinson's disease. Selective pulses of duration comparable the rotor period give optimal performance, but must be synchronized with the spinning in non-trivial ways to minimize chemical shift anisotropy recoupling effects. Soft pulses with a small bandwidth-duration product are best for exciting the ∼70 ppm bandwidth required for aliphatic-only dimensions.

Original languageEnglish (US)
Pages (from-to)245-257
Number of pages13
JournalJournal of Biomolecular NMR
Volume34
Issue number4
DOIs
StatePublished - Apr 2006

Keywords

  • CCC 3D
  • CSA
  • Dispersion
  • Resolution
  • Selective pulse
  • Solid-state NMR

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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