Summary
We have used a spin-echo difference NMR pulse sequence to measure three-bond J couplings between δ- and α-carbons of the leucine residues in a micelle-associated helical peptide dimer that corresponds to residues 62–101 of the transmembrane erythrocyte protein glycophorin A. The observed 3J couplings correlate strongly with the 13C chemical shift of the δ-methyl groups, and within experimental error both the shift distribution of the methyl carbons and the variations in 3J can be accounted for by variations in side-chain rotamer populations. We infer that all leucine side chains in this peptide dimer are in fast exchange among X 2 rotamers and sample two of the three possible rotameric states, even when the side chain forms part of the dimer interface. The observed correlation of chemical shift with couplings can be traced to a γ-gauche interaction of methyl and α-carbons. This correlation may provide an alternate route to rotamer analysis in some protein systems.
References
AlexandrescuA.T., AbeygunawardanaC. and ShortleD. (1994) Biochemistry, 33, 1063–1072.
ArcherS.J., IkuraM., TorchiaD.A. and BaxA. (1991) J. Magn. Reson., 95, 636–641.
BaxA., MaxD. and ZaxD. (1992) J. Am. Chem. Soc., 114, 6923–6925.
BoothH. (1964) Tetrahedron, 20, 2211–2216.
BystrovV.F. (1976) Prog. NMR Spectrosc., 10, 41–81.
DallingD.K. and GrantD.M. (1967) J. Am. Chem. Soc., 89, 6612–6622.
DavisA.L., KeelerJ., LaueE.D. and MoskauD. (1992) J. Magn. Reson., 98, 207–216.
EmsleyL. and BodenhausenG. (1990) Chem. Phys. Lett., 165, 469–476.
FarrowN.A., ZhangO., Forman-KayJ.D. and KayL.E. (1995) Biochemistry, 34, 868–875.
FrankM.K., CloreG.M. and GronenbornA.M. (1995) Protein Sci., 4, 2605–2615.
GriesingerC., SørensenO.W. and ErnstR.R. (1986) J. Chem. Phys., 85, 6837–6852.
GrzesiekS., VuisterG.W. and BaxA. (1993) J. Biomol. NMR, 3, 487–493.
KrivdinL.B. and DellaE.W. (1991) Prog. NMR Spectrosc., 23, 301–610.
LemmonM.A., FlanaganJ.M., TreutleinH.R., ZhangJ. and EngelmanD.M. (1992) Biochemistry, 31, 12719–12725.
LoganT.M., ThériaultY. and FesikS.W. (1994) J. Mol. Biol., 236, 637–648.
MontelioneG.T. and WagnerG. (1989) J. Am. Chem. Soc., 111, 5474–5475.
NeriD., BilleterM., WiderG. and WüthrichK. (1992) Science, 257, 1559–1563.
NicholsonL.K., KayL.E., BaldisseriD.M., ArangoJ., YoungP.E., BaxA. and TorchiaD.A. (1992) Biochemistry, 31, 5253–5263.
PardiA., BilleterM. and WüthrichK. (1984) J. Mol. Biol., 180, 741–751.
RexrothA., SchmidtP., SzalmaS., GeppertT., SchwalbeH. and GriesingerC. (1995) J. Am. Chem. Soc., 117, 10389–10390.
SchmiederP., ThanabalV., McIntoshL.P., DahlquistF.W. and WagnerG. (1991) J. Am. Chem. Soc., 113, 6323–6324.
ShakaA.J., BarkerP.B. and FreemanR. (1985) J. Magn. Reson., 64, 547–552.
ShortleD. and AbeygunawardanaC. (1993) Structure, 1, 121–134.
TolmanJ.R., ChungJ. and PrestegardJ.H. (1992) J. Magn. Reson., 98, 462–467.
VuisterG.W. and BaxA. (1993) J. Am. Chem. Soc., 115, 7772–7777.
VuisterG.W., WangA. and BaxA. (1993a) J. Am. Chem. Soc., 115, 5334–5335.
VuisterG.W., YamazakiT., TorchiaD.A. and BaxA. (1993b) J. Biomol. NMR, 3, 297–306.
VuisterG.W., GrzesiekS., DelaglioF., WangA.C., TschudinR., ZhuG. and BaxA. (1994) Methods Enzymol., 239, 79–105.
WagnerG. (1990) Prog. Magn. Reson., 22, 101–139.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
MacKenzie, K.R., Prestegard, J.H. & Engelman, D.M. Leucine side-chain rotamers in a glycophorin A transmembrane peptide as revealed by three-bond carbon—carbon couplings and 13C chemical shifts. J Biomol NMR 7, 256–260 (1996). https://doi.org/10.1007/BF00202043
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00202043