title: Mapping protein-protein interaction by 13C'-detected heteronuclear NMR spectroscopy.
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 (1)H-(15)N 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 (1)H-(15)N 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 (13)C direct-detection NMR is a useful complement to (1)H based NMR spectroscopy for monitoring protein-protein and protein-ligand interactions.
authors.affiliation: Department of Chemistry, University of Florence, 50019, Sesto Fiorentino, Italy. ivanobertini@cerm.unifi.it
authors.forename: Ivano
authors.initials: I
authors.lastname: Bertini
authors.forename: Isabella C
authors.initials: IC
authors.lastname: Felli
authors.forename: Leonardo
authors.initials: L
authors.lastname: Gonnelli
authors.forename: Roberta
authors.initials: R
authors.lastname: Pierattelli
authors.forename: Zinovia
authors.initials: Z
authors.lastname: Spyranti
authors.forename: Georgios A
authors.initials: GA
authors.lastname: Spyroulias
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry*; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D025941:Protein Interaction Mapping / Q000379:methods*; D017434:Protein Structure, Tertiary; D011522:Protons; D029701:Saccharomyces cerevisiae Proteins; D013329:Structure-Activity Relationship
chemical_list: D002247:Carbon Isotopes; D008024:Ligands; D011522:Protons; D029701:Saccharomyces cerevisiae Proteins