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Mapping protein–protein interaction by 13C′-detected heteronuclear NMR spectroscopy

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Abstract

The copper-mediated protein–protein interaction between yeast Atx1 and Ccc2 has been examined by protonless heteronuclear NMR and compared with the already available 1H–15N HSQC information. The observed chemical shift variations are analyzed with respect to the actual solution structure, available through intermolecular NOEs. The advantage of using the CON-IPAP spectrum with respect to the 1H–15N HSQC resides in the increased number of signals observed, including those of prolines. CBCACO-IPAP experiments allow us to focus on the interaction region and on side-chain carbonyls, while a newly designed CEN-IPAP experiment on side-chains of lysines. An attempt is made to rationalize the chemical shift variations on the basis of the structural data involving the interface between the proteins and the nearby regions. It is here proposed that protonless 13C direct-detection NMR is a useful complement to 1H based NMR spectroscopy for monitoring protein–protein and protein–ligand interactions.

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Abbreviations

δ:

chemical shift

IPAP:

in-phase-anti-phase

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Authors and Affiliations

  1. Department of Chemistry, University of Florence, 50019, Sesto Fiorentino, Italy

    Ivano Bertini, Isabella C. Felli & Roberta Pierattelli

  2. Magnetic Resonance Center (CERM), University of Florence, Via Luigi Sacconi 6, 50019, Sesto Fiorentino, Italy

    Ivano Bertini, Isabella C. Felli, Leonardo Gonnelli & Roberta Pierattelli

  3. Department of Pharmacy, University of Patras, 26504, Patras, Greece

    Zinovia Spyranti & Georgios A. Spyroulias

Authors
  1. Ivano Bertini
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  2. Isabella C. Felli
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  3. Leonardo Gonnelli
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  4. Roberta Pierattelli
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  5. Zinovia Spyranti
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  6. Georgios A. Spyroulias
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Correspondence to Ivano Bertini.

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Bertini, I., Felli, I.C., Gonnelli, L. et al. Mapping protein–protein interaction by 13C′-detected heteronuclear NMR spectroscopy. J Biomol NMR 36, 111–122 (2006). https://doi.org/10.1007/s10858-006-9068-z

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  • DOI: https://doi.org/10.1007/s10858-006-9068-z

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