Summary
We have developed an improved isotope-filtered pulse scheme in combination with a double-tuned filter, a hyperbolic secant inversion pulse, and a z-filter with a pulsed field gradient. These filtering pulse schemes have been incorporated into several one-, two-, and three-dimensional experiments, which were applied to the 13C/15N uniformly labeled N-terminal SH3 domain of Grb2 complexed with the unlabeled Sos-derived peptide. The proton resonances of the Sos-derived peptide were unambiguously assigned using isotope-filtered DQF-COSY, TOCSY and NOESY spectra. Furthermore, in the isotope-filtered, isotope-edited 3D NOESY spectrum, intermolecular NOEs between the labeled protein and the unlabeled peptide could be identified. Through these applications, we demonstrate the high filtering efficiency of the presented pulse scheme.
References
BaxA. and PochapskyS.S. (1992) J. Magn. Reson., 99, 638–643.
BodenhausenG. and RubenD.J. (1980) Chem. Phys. Lett. 69, 185–189.
BurgeringM.J.M., BoelensR., CaffreyM., BregJ.N. and KapteinR. (1993a) FEBS Lett. 330, 105–109.
BurgeringM.J.M., BoelensR. and KapteinR. (1993b) J. Biomol. NMR, 3, 709–714.
FesikS.W., GampeJr.R.T., EatonH.L., GemmeckerG., OlejniczakE.T., NeriP., HoltzmanT.F., EganD.A., EdaljiR., SimmerR., HelfrichR., HochlowskiJ. and JacksonM. (1991) Biochemistry 30, 6574–6583.
FolkersP.J.M., FolmerR.H.A., KoningsR.N.H. and HilbersC.W. (1993) J. Am. Chem. Soc., 115, 3798–3799.
FolmerR.H.A., HilbersC.W., KoningsR.N.H. and HallengaK. (1995) J. Biomol. NMR, 5, 427–432.
GeenH. and FreemanR. (1991) J. Magn. Reson., 93, 93–141.
GemmeckerG., OlejniczakE.T. and FesikS.W. (1992) J. Magn. Reson., 96, 199–204.
GriesingerC., OttingG., WüthrichK. and ErnstR.R. (1988) J. Am. Chem. Soc., 110, 7870–7872.
GrzesiekS. and BaxA. (1993) J. Biomol. NMR, 3, 185–204.
HallengaK. and LippensG.M. (1995) J. Biomol. NMR, 5, 59–66.
IkuraM. and BaxA. (1992) J. Am. Chem. Soc., 114, 2433–2440.
IkuraM., CloreG.M., GronenbornA.M., ZhuG., KleeC.B. and BaxA. (1992) Science, 256, 632–638.
LeeW., HarveyT.S., YinY., YauP., LitchfieldD. and ArrowsmithC.H. (1994a) Nat. Struct. Biol. 1, 877–890.
LeeW., RevingtonM.J., ArrowsmithC. and KayL.E. (1994b) FEBS Lett., 350, 87–90.
MüllerL. (1979) J. Am. Chem. Soc., 101, 4481–4484.
OguraK., TerasawaH. and InagakiF. (1996) J. Magn. Reson., B112, 63–68.
OttingG., SennH., WagnerG. and WüthrichK. (1986) J. Magn. Reson., 70, 500–505.
OttingG. and WüthrichK. (1989) J. Magn. Reson., 85, 586–594.
PetrosA.M., KawaiM., LulyJ.R. and FesikS.W. (1992) FEBS Lett., 308, 309–314.
RosenN. and ZenerC. (1932) Phys. Rev., 40, 502–507.
SilverM.S., JosephR.I. and HoultD.I. (1984a) J. Magn. Reson., 59, 347–353.
SilverM.S., JosephR.I. and HoultD.I. (1984b) Phys. Rev., A31, 2753–2755.
SmallcombeS.H. (1993) J. Am. Chem. Soc., 115, 4776–4785.
TerasawaH., KohdaD., HatanakaH., TsuchiyaS., OguraK., NagataK., IshiiS., MandiyanV., UllrichA., SchlessingerJ. and InagakiF. (1994) Nat. Struct. Biol., 1,891–897.
WeberC., WiderG., VonFreybergB., TraberR., BraunW., WidmerH. and WüthrichK. (1991) Biochemistry, 30, 6563–6574.
ZhangH., ZhaoD., RevingtonM., LeeW., JiaX., ArrowsmithC. and JardetskyO. (1994) J. Mol. Biol., 238, 592–614.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ogura, K., Terasawa, H. & Inagaki, F. An improved double-tuned and isotope-filtered pulse scheme based on a pulsed field gradient and a wide-band inversion shaped pulse. J Biomol NMR 8, 492–498 (1996). https://doi.org/10.1007/BF00228150
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00228150