Towards automatic protein backbone assignment using proton-detected 4D solid-state NMR data

J Biomol NMR. 2014 Nov;60(2-3):85-90. doi: 10.1007/s10858-014-9859-6. Epub 2014 Sep 6.

Abstract

We introduce an efficient approach for sequential protein backbone assignment based on two complementary proton-detected 4D solid-state NMR experiments that correlate Hi(N)/Ni with CAi/COi or CAi-1/COi-1. The resulting 4D spectra exhibit excellent sensitivity and resolution and are amenable to (semi-)automatic assignment approaches. This strategy allows to obtain sequential connections with high confidence as problems related to peak overlap and multiple assignment possibilities are avoided. Non-uniform sampling schemes were implemented to allow for the acquisition of 4D spectra within a few days. Rather moderate hardware requirements enable the successful demonstration of the method on deuterated type III secretion needles using a 600 MHz spectrometer at a spinning rate of 25 kHz.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation
  • Bacterial Proteins / chemistry*
  • Proton Magnetic Resonance Spectroscopy*
  • Salmonella typhimurium / metabolism

Substances

  • Bacterial Proteins