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Journal of Biomolecular NMR

, Volume 15, Issue 2, pp 151–155 | Cite as

A TROSY CPMG sequence for characterizing chemical exchange in large proteins

  • J. Patrick Loria
  • Mark Rance
  • Arthur G. PalmerIII
Article

Abstract

A new NMR spin relaxation experiment is described for measuring chemical exchange time constants from approximately 0.5 ms to 5 ms in 15N-labeled macromolecules. The pulse sequence is based on the Carr–Purcell–Meiboom–Gill technique [Carr and Purcell (1954) Phys. Rev., 94, 630–638; Meiboom and Gill (1958) Rev. Sci. Instrum., 29, 688–691; Loria et al. (1999) J. Am. Chem. Soc., 121, 2331–2332], but implements TROSY selection [Pervushin et al. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366–12371] to permit measurement of exchange linebroadening contributions to the narrower component of the 1H-15N scalar-coupled doublet. This modification extends the size limitation imposed on relaxation measurements due to the fast decay of transverse magnetization in larger macromolecules. The new TROSY-CPMG experiment is demonstrated on a [U-98% 15 N] labeled sample of basic pancreatic trypsin inhibitor and a [U-83% 2H, U-98% 15 N] labeled sample of triosephosphate isomerase, a 54 kDa homodimeric protein.

BPTI chemical exchange CPMG triosephosphate isomerase TROSY 

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Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • J. Patrick Loria
    • 1
  • Mark Rance
    • 2
  • Arthur G. PalmerIII
    • 1
  1. 1.Department of Biochemistry and Molecular BiophysicsColumbia UniversityNew YorkU.S.A.
  2. 2.Department of Molecular Genetics, Biochemistry and MicrobiologyUniversity of Cincinnati College of MedicineCincinnatiU.S.A.

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