Abstract
Water molecules are a major determinant of protein stability and are important for understanding protein–protein interactions. We present two experiments which allow to measure first the effective T2 decay rate of individual amide proton, and second the magnetization build-up rates for a selective transfer from H2O to HN using spin diffusion as a mixing element. The experiments are demonstrated for a uniformly 2H, 15N labeled sample of a microcrystalline SH3 domain in which exchangeable deuterons were back-substituted with protons. In order to evaluate the NMR experimental data, as X-ray structure of the protein was determined using the same crystallization protocol as for the solid-state NMR sample. The NMR experimental data are correlated with the dipolar couplings calculated from H2O–HN distances which were extracted from the X-ray structure of the protein. We find that the HN T2 decay rates and H2O–HN build-up rates are sensitive to distance and dynamics of the detected water molecules with respect to the protein. We show that qualitative information about localization and dynamics of internal water molecules can be obtained in the solid-state by interpretation of the spin dynamics of a reporter amide proton.
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M. Bak J.T. Rasmussen N.C. Nielsen (2000) J. Magn. Reson. 147 296–330 Occurrence Handle10.1006/jmre.2000.2179 Occurrence Handle1:CAS:528:DC%2BD3cXotlKlurk%3D Occurrence Handle11097821
A.E. Bennett C.M. Rienstra M. Auger K.V. Lakshmi R.G. Griffin (1995) J. Chem. Phys. 103 6951–6958 Occurrence Handle10.1063/1.470372 Occurrence Handle1:CAS:528:DyaK2MXovVeqtLs%3D
S. Cai S.Y. Stevens A.P. Budor E.R.P. Zuiderweg (2003) Biochemistry 42 11100–11108 Occurrence Handle1:CAS:528:DC%2BD3sXnt1Ohs7o%3D Occurrence Handle14503860
F. Castellani B.-J. Rossum Particlevan A. Diehl M. Schubert K. Rehbein H. Oschkinat (2002) Nature 420 98–102 Occurrence Handle1:CAS:528:DC%2BD38XosVCmurk%3D Occurrence Handle12422222
V. Chevelkov B.J.V. Rossum F. Castellani K. Rehbein A. Diehl M. Hohwy S. Steuernagel F. Engelke H. Oschkinat B. Reif (2003) J. Am. Chem. Soc. 125 7788–7789 Occurrence Handle1:CAS:528:DC%2BD3sXktlGqsro%3D Occurrence Handle12822982
G.M. Clore A. Bax P.T. Wingfield A.M. Gronenborn (1990) Biochemistry 29 5671–5676 Occurrence Handle1:CAS:528:DyaK3cXktVCgtbY%3D Occurrence Handle2383553
V.P. Denisov J. Peters H.D. Hörlein B. Halle (1996) Nat. Struct. Biol. 3 505–510 Occurrence Handle1:CAS:528:DyaK28Xjtlymu7s%3D Occurrence Handle8646535
G. Gemmecker W. Jahnke H. Kessler (1993) J. Am. Chem. Soc. 115 11620–11621 Occurrence Handle1:CAS:528:DyaK3sXms1CqsLs%3D
M. Gottschalk N.A. Dencher B. Halle (2001) J. Mol. Biol. 311 605–621 Occurrence Handle1:CAS:528:DC%2BD3MXlslGitLc%3D Occurrence Handle11493013
B. Halle V.P. Denisov (2001) Meth. Enzymol. 338 178–201 Occurrence Handle1:CAS:528:DC%2BD3MXls1elt7w%3D Occurrence Handle11460548
G.S. Harbison J.E. Roberts J. Herzfeld R.G. Griffin (1988) J. Am. Chem. Soc. 110 7221–7223 Occurrence Handle1:CAS:528:DyaL1cXlsVGlsL4%3D
W.J. Kabsch (1993) J. Appl. Cryst. 26 795–800 Occurrence Handle1:CAS:528:DyaK2cXptFeltw%3D%3D
R. Koradi M. Billeter K. Wüthrich (1996) J. Mol. Graph. 14 51–55 Occurrence Handle1:CAS:528:DyaK28Xis12nsrc%3D Occurrence Handle8744573
A. Lesage A. Böckmann (2003) J. Am. Chem. Soc. 125 13336–13337 Occurrence Handle1:CAS:528:DC%2BD3sXnvFOisLw%3D Occurrence Handle14583011
A. Lesage L. Emsley (2001) J. Magn. Reson. 148 449–454 Occurrence Handle1:CAS:528:DC%2BD3MXhsFOgsLo%3D Occurrence Handle11237652
M.H. Levitt D.P. Raleigh F. Creuzet R.G. Griffin (1990) J. Chem. Phys. 92 6347–6364 Occurrence Handle1:CAS:528:DyaK3cXks12rt7g%3D
A. McDermott T. Polenova A. Böckmann K.W. Zilm E.K. Paulsen R.W. Martin G.T. Montelione (2000) J. Biomol. NMR 16 209–219 Occurrence Handle1:CAS:528:DC%2BD3cXivFKrs7c%3D Occurrence Handle10805127
K. Murata K. Mitsuoka T. Hirai T. Walz P. Agre J.B. Heymann A. Engel Y. Fujiyoshi (2000) Nature 407 599–605 Occurrence Handle10.1038/35036519 Occurrence Handle1:CAS:528:DC%2BD3cXnsVKlsLg%3D Occurrence Handle11034202
G.N. Murshudov A.A. Vagin E.J. Dodson (1997) Acta Cryst. D53 240–255 Occurrence Handle1:CAS:528:DyaK2sXjs1Gnsb4%3D
A. Musacchio M.E.M. Noble R. Pauptit R.K. Wierenga M. Saraste (1992) Nature 359 851–855 Occurrence Handle1:CAS:528:DyaK38XmsVOgu7w%3D Occurrence Handle1279434
J. Navaza (1994) Acta Cryst. A50 157–163 Occurrence Handle1:CAS:528:DyaK2cXitlaksbk%3D
F. Noack (1986) Prog. NMR Spect. 18 171–276 Occurrence Handle1:CAS:528:DyaL28XhvFynurk%3D
F. Noack S. Becker J. Struppe (1997) Annu. Rep. NMR Spectrosc. 33 1 Occurrence Handle1:CAS:528:DyaK2sXktFahsLk%3D
G. Otting (1997) Prog. Nucl. Magn. Reson. Spectrosc. 31 259–285 Occurrence Handle1:CAS:528:DyaK1cXnsFGntg%3D%3D
G. Otting K. Wüthrich (1989) J. Am. Chem. Soc. 111 1871–1875 Occurrence Handle1:CAS:528:DyaL1MXhtFSgsLo%3D
J. Pauli M. Baldus B.-J. Van Rossum H. De Groot H. Oschkinat (2001) Chem. BioChem. 2 272–281 Occurrence Handle1:CAS:528:DC%2BD3MXivVCntbY%3D
J. Pauli B.-J. Van Rossum H. Förster H.J.M. De Groot H. Oschkinat (2000) J. Magn. Reson. 143 411–416 Occurrence Handle1:CAS:528:DC%2BD3cXhvVyjtr4%3D Occurrence Handle10729269
E.K. Paulson C.R. Morcombe V. Gaponenko B. Dancheck R.A. Byrd K.W. Zilm (2003) J. Am. Chem. Soc. 125 14222–14223 Occurrence Handle1:CAS:528:DC%2BD3sXos1Whsbw%3D Occurrence Handle14624539
E.K. Paulson C.R. Morcombe V. Gaponenko B. Dancheck R.A. Byrd K.W. Zilm (2003) J. Am. Chem. Soc. 125 15831–15836 Occurrence Handle1:CAS:528:DC%2BD3sXptlOmurg%3D Occurrence Handle14677974
B. Reif R.G. Griffin (2003) J. Magn. Reson. 160 78–83 Occurrence Handle1:CAS:528:DC%2BD3sXmslejsQ%3D%3D Occurrence Handle12565053
B. Reif C.P. Jaroniec C.M. Rienstra M. Hohwy R.G. Griffin (2001) J. Magn. Reson. 151 320–327 Occurrence Handle1:CAS:528:DC%2BD3MXmsVGisLg%3D Occurrence Handle11531354
B. Reif B.J. Rossum Particlevan F. Castellani K. Rehbein A. Diehl H. Oschkinat (2003) J. Am. Chem. Soc. 125 1488–1489 Occurrence Handle1:CAS:528:DC%2BD3sXksFektg%3D%3D Occurrence Handle12568603
C.M. Rienstra L. Tucker-Kellogg C.P. Jaroniec M. Hohwy B. Reif M.T. McMahon B. Tidor T. Lozano-Pérez R.G. Griffin (2002) Proc. Natl. Acad. Sci. USA 99 10260–10265 Occurrence Handle1:CAS:528:DC%2BD38Xmt1CitLY%3D Occurrence Handle12149447
M.K. Rosen K.H. Gardner R.C. Willis W.E. Parris T. Pawson L.E. Kay (1996) J. Mol. Biol. 263 627–636 Occurrence Handle1:CAS:528:DyaK28XntFyqs7c%3D Occurrence Handle8947563
S.M. Saparov P. Pohl (2004) Proc. Natl. Acad. Sci. USA 101 4805–4809 Occurrence Handle1:CAS:528:DC%2BD2cXjsFCjuro%3D Occurrence Handle15034178
B.-J. Rossum Particlevan F. Castellani J. Pauli K. Rehbein J. Hollander H.J.M. De Groot H. Oschkinat (2003) J. Biomol. NMR 25 217–223 Occurrence Handle12652133
M.C. Vega J.C. Martinez L. Serrano (2000) Prot. Sci. 9 2322–2328 Occurrence Handle1:CAS:528:DC%2BD3MXns1aksA%3D%3D
E. Vinogradov P.K. Madhu S. Vega (1999) Chem. Phys. Lett. 314 443–450 Occurrence Handle1:CAS:528:DyaK1MXotVKrtr0%3D
D. Voet J.G. Voet (1995) Biochemistry, 2nd edn John Wiley and Sons New York
Y. Zhou J.H. Morais-Cabral A. Kaufman R. MacKinnon (2001) Nature 414 43–48 Occurrence Handle1:CAS:528:DC%2BD3MXot1Wqsbg%3D Occurrence Handle11689936
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Chevelkov, V., Faelber, K., Diehl, A. et al. Detection of dynamic water molecules in a microcrystalline sample of the SH3 domain of α-spectrin by MAS solid-state NMR. J Biomol NMR 31, 295–310 (2005). https://doi.org/10.1007/s10858-005-1718-z
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DOI: https://doi.org/10.1007/s10858-005-1718-z