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Measurement of slow (μs-ms) time scale dynamics in protein side chains by 15N relaxation dispersion NMR spectroscopy: Application to Asn and Gln residues in a cavity mutant of T4 lysozyme

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Measurement of slow (μs-ms) time scale dynamics in protein side chains by 15N relaxation dispersion NMR spectroscopy : Application to Asn and Gln residues in a cavity mutant of T4 lysozyme. / Mulder, F. A A; Skrynnikov, N. R.; Hon, B. et al.

In: Journal of the American Chemical Society, Vol. 123, No. 5, 07.02.2001, p. 967-975.

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@article{461092eedf9d41518b4ad79ce0ec2d20,
title = "Measurement of slow (μs-ms) time scale dynamics in protein side chains by 15N relaxation dispersion NMR spectroscopy: Application to Asn and Gln residues in a cavity mutant of T4 lysozyme",
abstract = "A new NMR experiment is presented for the measurement of μs-ms time scale dynamics of Asn and Gln side chains in proteins. Exchange contributions to the 15N line widths of side chain residues are determined via a relaxation dispersion experiment in which the effective nitrogen transverse relaxation rate is measured as a function of the number of refocusing pulses in constant-time, variable spacing CPMG intervals. The evolution of magnetization from scalar couplings and dipole-dipole cross-correlations, which has limited studies of exchange in multi-spin systems in the past, does not affect the extraction of accurate exchange parameters from relaxation profiles of NH2 groups obtained in the present experiment. The utility of the method is demonstrated with an application to a Leu → Ala cavity mutant of T4 lysozyme, L99A. It is shown that many of the side chain amide groups of Asn and Gln residues in the C-terminal domain of the protein are affected by a chemical exchange process which may be important in facilitating the rapid binding of hydrophobic ligands to the cavity.",
author = "Mulder, {F. A A} and Skrynnikov, {N. R.} and B. Hon and Dahlquist, {F. W.} and Kay, {L. E.}",
year = "2001",
month = feb,
day = "7",
doi = "10.1021/ja003447g",
language = "English",
volume = "123",
pages = "967--975",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "ACS Publications",
number = "5",

}

RIS

TY - JOUR

T1 - Measurement of slow (μs-ms) time scale dynamics in protein side chains by 15N relaxation dispersion NMR spectroscopy

T2 - Application to Asn and Gln residues in a cavity mutant of T4 lysozyme

AU - Mulder, F. A A

AU - Skrynnikov, N. R.

AU - Hon, B.

AU - Dahlquist, F. W.

AU - Kay, L. E.

PY - 2001/2/7

Y1 - 2001/2/7

N2 - A new NMR experiment is presented for the measurement of μs-ms time scale dynamics of Asn and Gln side chains in proteins. Exchange contributions to the 15N line widths of side chain residues are determined via a relaxation dispersion experiment in which the effective nitrogen transverse relaxation rate is measured as a function of the number of refocusing pulses in constant-time, variable spacing CPMG intervals. The evolution of magnetization from scalar couplings and dipole-dipole cross-correlations, which has limited studies of exchange in multi-spin systems in the past, does not affect the extraction of accurate exchange parameters from relaxation profiles of NH2 groups obtained in the present experiment. The utility of the method is demonstrated with an application to a Leu → Ala cavity mutant of T4 lysozyme, L99A. It is shown that many of the side chain amide groups of Asn and Gln residues in the C-terminal domain of the protein are affected by a chemical exchange process which may be important in facilitating the rapid binding of hydrophobic ligands to the cavity.

AB - A new NMR experiment is presented for the measurement of μs-ms time scale dynamics of Asn and Gln side chains in proteins. Exchange contributions to the 15N line widths of side chain residues are determined via a relaxation dispersion experiment in which the effective nitrogen transverse relaxation rate is measured as a function of the number of refocusing pulses in constant-time, variable spacing CPMG intervals. The evolution of magnetization from scalar couplings and dipole-dipole cross-correlations, which has limited studies of exchange in multi-spin systems in the past, does not affect the extraction of accurate exchange parameters from relaxation profiles of NH2 groups obtained in the present experiment. The utility of the method is demonstrated with an application to a Leu → Ala cavity mutant of T4 lysozyme, L99A. It is shown that many of the side chain amide groups of Asn and Gln residues in the C-terminal domain of the protein are affected by a chemical exchange process which may be important in facilitating the rapid binding of hydrophobic ligands to the cavity.

U2 - 10.1021/ja003447g

DO - 10.1021/ja003447g

M3 - Journal article

C2 - 11456632

AN - SCOPUS:0035819455

VL - 123

SP - 967

EP - 975

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 5

ER -