A TROSY CPMG sequence for characterizing chemical exchange in large proteins

Journal of Biomolecular NMR - Tập 15 - Trang 151-155 - 1999
J. Patrick Loria1, Mark Rance2, Arthur G. Palmer1
1Department of Biochemistry and Molecular Biophysics, Columbia University, New York, U.S.A.
2Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, U.S.A.

Tóm tắt

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.

Tài liệu tham khảo

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