De novo determination of peptide structure with solid-state magic-angle spinning NMR spectroscopy

doi:10.1073/pnas.152346599

De novo determination of peptide structure with solid-state magic-angle spinning NMR spectroscopy

^*Department of Chemistry, ^†Francis Bitter Magnet Laboratory, ^§Department of Electrical Engineering and Computer Science, and ^**Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA 02139

Communicated by John S. Waugh, Massachusetts Institute of Technology, Cambridge, MA (received for review January 25, 2002)

Abstract

The three-dimensional structure of the chemotactic peptide N-formyl-l-Met-l-Leu-l-Phe-OH was determined by using solid-state NMR (SSNMR). The set of SSNMR data consisted of 16 ¹³C–¹⁵N distances and 18 torsion angle constraints (on 10 angles), recorded from uniformly ¹³C,¹⁵N- and ¹⁵N-labeled samples. The peptide's structure was calculated by means of simulated annealing and a newly developed protocol that ensures that all of conformational space, consistent with the structural constraints, is searched completely. The result is a high-quality structure of a molecule that has thus far not been amenable to single-crystal diffraction studies. The extensions of the SSNMR techniques and computational methods to larger systems appear promising.

Footnotes

↵ ‡ Present address: Department of Chemistry, University of Illinois, Urbana, IL 61801.
↵ ¶ Present address: Laboratory for Physical Chemistry, ETH-Hönggerberg, CH-8093 Zürich, Switzerland.
↵ ∥ Present address: Laboratory for Organic Chemistry, TU-Munich, D-85647 Munich, Germany.
↵ †† To whom reprint requests may be addressed. E-mail: rgg{at}mit.edu, tlp{at}mit.edu, or tidor{at}mit.edu.
Abbreviations:
1. SSNMR, solid-state NMR
2. rmsd, root-mean-square deviation
3. MAS, magic-angle spinning
4. f-MLF-OH, N-formyl-l-Met-l-Leu-l-Phe-OH
5. f-MLF-OMe, f-MLF-OH methyl ester
6. 2D and 3D, two- and three-dimensional
7. REDOR, rotational-echo double-resonance