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Assignment of the protonated 13C resonances of apo-neocarzinostatin by 2D heteronuclear NMR spectroscopy at natural abundance

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Summary

Nearly complete assignment of the protonated carbon resonances of apo-neocarzinostatin, 113-amino acid antitumor antibiotic carrier protein, has been achieved at natural 13C abundance using heteronuclear 2D experiments. Most of the cross peaks in the proton-carbon correlation map were identified by the combined use of HMQC, HMQC-RELAY and HMQC-NOESY spectra, using already published proton chemical shifts. However, double-DEPT and triple-quantum experiments had to be performed for the edition of CH and CH2 side-chain groups, respectively, which were hardly visible on HMQC-type maps. The triple-quantum pulse sequence was adapted from its original scheme to be applicable to a natural abundance sample. The correlation between carbon chemical shifts and the apo-neocarzinostatin structure is discussed. In particular, 13C alpha secondary shifts correlate well with the backbone conformation. These shifts also yield information about the main-chain flexibility of the protein. Assignments reported herein will be used further for interpretation of carbon relaxation times in a study of the internal dynamics of apo-neocarzinostatin.

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References

  • AdjadjE., MispelterJ., QuiniouE., DimicoliJ.L., FavaudonV. and LhosteJ.M. (1990) Eur. J. Biochem., 190, 263–271.

    PubMed  Google Scholar 

  • AdjadjE., QuiniouE., MispelterJ., FavaudonV. and LhosteJ.M. (1992a) Eur. J. Biochem., 203, 505–511.

    PubMed  Google Scholar 

  • AdjadjE., QuiniouE., MispelterJ., FavaudonV. and LhosteJ.M. (1992b) Biochimie, 74, 853–858.

    Article  PubMed  Google Scholar 

  • BaxA., GriffeyR.H. and HawkinsB.L. (1983) J. Magn. Reson., 55, 301–315.

    Google Scholar 

  • BaxA., IkuraM., KayL.E., TorchiaD.A. and TschudinR. (1990) J. Magn. Reson., 86, 304–318.

    Google Scholar 

  • BendallM.R., PeggD.T. and DoddrellD.M. (1983) J. Magn. Reson., 52, 81–117.

    Google Scholar 

  • BodenhausenG. and RubenD.J. (1980) Chem. Phys. Lett., 69, 185–189.

    Article  Google Scholar 

  • BrühwilerD. and WagnerG. (1986) J. Magn. Reson., 69, 546–551.

    Google Scholar 

  • CloreG.M., BaxA., DriscollP.C., WingfieldP.T. and GronenbornA.M. (1990) Biochemistry, 29, 8172–8184.

    PubMed  Google Scholar 

  • DavisD.G. (1990) J. Magn. Reson., 90, 589–596.

    Google Scholar 

  • DavisD.G. (1991) J. Magn. Reson., 91, 665–672.

    Google Scholar 

  • deDiosA.C., PearsonJ.G. and OldfieldE. (1993) Science, 260, 1491–1496.

    PubMed  Google Scholar 

  • DoddrellD.M., PeggD.T. and BendallM.R. (1982) J. Magn. Reson., 48, 323–327.

    Google Scholar 

  • DomkeT. and LeibfritzD. (1990) J. Magn. Reson., 88, 401–405.

    Google Scholar 

  • ErnstR.R., BodenhausenG. and WokaumA. (1987) In International Series of Monographs on Chemistry, Vol. 14 (Eds, BreslowR., HalpernJ. and RowlinsonJ.S.) Clarendon Press, Oxford, pp. 467–489.

    Google Scholar 

  • GaoX. and BurkhartW. (1991) Biochemistry, 30, 7730–7739.

    PubMed  Google Scholar 

  • GaoX. (1992) J. Mol. Biol., 225, 125–135.

    PubMed  Google Scholar 

  • HowarthO.W. and LilleyD.M.J. (1978) Prog. NMR Spectrosc., 12, 1–40.

    Google Scholar 

  • IkuraM., SperaS., BarbatoG., KayL.E., KrinksM. and BaxA. (1991) Biochemistry, 30, 9216–9228.

    PubMed  Google Scholar 

  • KappenL.S., NapierM.A. and GoldbergI.H. (1980) Proc. Natl. Acad. Sci. USA, 77, 1970–1974.

    PubMed  Google Scholar 

  • KayL.E., JueT.L., BangerterB. and DemouP.C. (1987) J. Magn. Reson., 73, 558–564.

    Google Scholar 

  • KayL.E. and BaxA. (1989) J. Magn. Reson., 84, 598–603.

    Google Scholar 

  • KesslerH., SchmiederP., and KurzM. (1989) J. Magn. Reson., 85, 400–405.

    Google Scholar 

  • KimK.H., KwonB.M., MyersA.G. and ReesD.C. (1993) Science, 262, 1042–1046.

    PubMed  Google Scholar 

  • LawsD.D., deDiosA.C. and OldfieldE. (1993) J. Biomol. NMR, 3, 607–612.

    Article  PubMed  Google Scholar 

  • LernerL. and BaxA. (1986) J. Magn. Reson., 69, 375–380.

    Google Scholar 

  • LondonR.E. and AvitabileJ. (1978) J. Am. Chem. Soc., 100, 7159–7165.

    Google Scholar 

  • MüllerL. (1979) J. Am. Chem. Soc., 101, 4481–4484.

    Google Scholar 

  • NirmalaN.R. and WagnerG. (1988) J. Am. Chem. Soc., 110, 7557–7558.

    Google Scholar 

  • NortonR.S., ClouseA.O., AddlemanR. and AllerhandA. (1977) J. Am. Chem. Soc., 99, 79–83.

    PubMed  Google Scholar 

  • NorwoodT.J., BoydJ., HeritageJ.E., SoffeN. and CampbellI.D. (1990) J. Magn. Reson., 87, 488–501.

    Google Scholar 

  • OhB.H., WestlerW.M. and MarkleyJ.L. (1989) J. Am. Chem. Soc., 111, 3083–3085.

    Google Scholar 

  • OldfieldE., NortonR.S. and AllerhandA. (1975) J. Biol. Chem., 250, 6368–6380.

    PubMed  Google Scholar 

  • PalmerIIIA.G., CavanaghJ., WrightP.E. and RanceM. (1991) J. Magn. Reson., 93, 151–170.

    Google Scholar 

  • PastoreA. and SaudekV. (1990) J. Magn. Reson., 90, 165–176.

    Google Scholar 

  • RemerowskiM.L., GlaserS.J., SiekerL.C., SamyT.S.A. and DrobnyG.P. (1990) Biochemistry, 29, 8401–8409.

    PubMed  Google Scholar 

  • RibeiroA.A., KingR., RestivoC. and JardetzkyO. (1980) J. Am. Chem. Soc., 102, 4040–4051.

    Google Scholar 

  • RicharzR., NagayamaK. and WüthrichK. (1980) Biochemistry, 19, 5189–5196.

    PubMed  Google Scholar 

  • SchmidtJ.M. and RüterjansH. (1990) J. Am. Chem. Soc., 112, 1279–1280.

    Google Scholar 

  • ShakaA.J., KeelerJ., FrenkielT. and FreemanR. (1983) J. Magn. Reson., 52, 335–338.

    Google Scholar 

  • ShonK. and OpellaS.J. (1989) J. Magn. Reson., 82, 193–197.

    Google Scholar 

  • SklenarV., TorchiaD. and BaxA. (1987) J. Magn. Reson., 73, 375–379.

    Google Scholar 

  • SørensenO.W., EichG.W., LevittM.H., BodenhausenG. and ErnstR.R. (1983) Prog. NMR Spectrosc., 16, 163–192.

    Google Scholar 

  • SperaS. and BaxA. (1991) J. Am. Chem. Soc., 113, 5490–5492.

    Google Scholar 

  • SummersM.F., MarzilliL.G. and BaxA. (1986) J. Am. Chem. Soc., 108, 4285–4294.

    Google Scholar 

  • TateS.I., MasuiY. and InagakiF. (1991) J. Magn. Reson., 94, 625–630.

    Google Scholar 

  • TeplyakovA., ObmolovaG., WilsonK. and KuromizuK. (1993) Eur. J. Biochem., 213, 737–741.

    PubMed  Google Scholar 

  • WagnerG. and BrühwilerD. (1986) Biochemistry, 25, 5839–5843.

    PubMed  Google Scholar 

  • WagnerG. (1989) Methods Enzymol., 176, 93–113.

    PubMed  Google Scholar 

  • WishartD.S., SykesB.D. and RichardsF.M. (1991) J. Mol. Biol., 222, 311–333.

    PubMed  Google Scholar 

  • WüthrichK. (1976) NMR in Biological Research: Peptides and Proteins, Elsevier, New York, NY, pp. 157–209 and 293–316.

    Google Scholar 

  • ZhangX. and WangC. (1991) J. Magn. Reson., 91, 618–623.

    Google Scholar 

  • ZuiderwegE.R.P. (1990) J. Magn. Reson., 86, 346–357.

    Google Scholar 

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Authors and Affiliations

  1. U350 INSERM, Institut Curie Biologie, Centre Universitaire, Bât. 112, F-91405, Orsay Cedex, France

    Claudine Lefevre, Élisabeth Adjadj, Éric Quiniou & Joël Mispelter

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  1. Claudine Lefevre
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  2. Élisabeth Adjadj
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  3. Éric Quiniou
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  4. Joël Mispelter
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Lefevre, C., Adjadj, É., Quiniou, É. et al. Assignment of the protonated 13C resonances of apo-neocarzinostatin by 2D heteronuclear NMR spectroscopy at natural abundance. J Biomol NMR 4, 689–702 (1994). https://doi.org/10.1007/BF00404278

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  • DOI: https://doi.org/10.1007/BF00404278

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