Journal of Biomolecular NMR

, Volume 16, Issue 3, pp 209–219 | Cite as

Partial NMR assignments for uniformly (13C, 15N)-enriched BPTI in the solid state

  • Ann McDermott
  • Tatyana Polenova
  • Anja Bockmann
  • Kurt.W. Zilm
  • Eric K. Paulsen
  • Rachel W. Martin
  • Gaetano T. Montelione

Abstract

We demonstrate that high-resolution multidimensional solid state NMR methods can be used to correlate many backbone and side chain chemical shifts for hydrated micro-crystalline U-13C,15N Basic Pancreatic Trypsin Inhibitor (BPTI), using a field strength of 800 MHz for protons, magic angle sample spinning rates of 20 kHz and proton decoupling field strengths of 140 kHz. Results from two homonuclear transfer methods, radio frequency driven dipolar recoupling and spin diffusion, were compared. Typical 13C peak line widths are 0.5 ppm, resulting in Cα-Cβ and Cα-CO regions that exhibit many resolved peaks. Two-dimensional carbon–carbon correlation spectra of BPTI have sufficient resolution to identify and correlate many of the spin systems associated with the amino acids. As a result, we have been able to assign a large number of the spin systems in this protein. The agreement between shifts measured in the solid state and those in solution is typically very good, although some shifts near the ion binding sites differ by at least 1.5 ppm. These studies were conducted with approximately 0.2 to 0.4 μmol of enriched material; the sensitivity of this method is apparently adequate for other biological systems as well.

amino acid side chain assignment protocol bovine pancreatic trypsin inhibitor solid state NMR uniform labeling 

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Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Ann McDermott
    • 1
  • Tatyana Polenova
    • 1
  • Anja Bockmann
    • 1
  • Kurt.W. Zilm
    • 2
  • Eric K. Paulsen
    • 2
  • Rachel W. Martin
    • 2
  • Gaetano T. Montelione
    • 3
  1. 1.Department of ChemistryColumbia UniversityNew YorkU.S.A.
  2. 2.Department of ChemistryYale UniversityNew HavenU.S.A.
  3. 3.Center for Advanced Biotechnology and MedicineRutgers UniversityPiscatawayU.S.A.

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