| First Author | Lindfors HE | Year | 2008 |
| Journal | J Biomol NMR | Volume | 41 |
| Issue | 3 | Pages | 157-67 |
| PubMed ID | 18560762 | Mgi Jnum | J:200397 |
| Mgi Id | MGI:5508591 | Doi | 10.1007/s10858-008-9248-0 |
| Citation | Lindfors HE, et al. (2008) Mobility of TOAC spin-labelled peptides binding to the Src SH3 domain studied by paramagnetic NMR. J Biomol NMR 41(3):157-67 |
| abstractText | Paramagnetic relaxation enhancement provides a tool for studying the dynamics as well as the structure of macromolecular complexes. The application of side-chain coupled spin-labels is limited by the mobility of the free radical. The cyclic, rigid amino acid spin-label TOAC (2,2,6,6-Tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid), which can be incorporated straightforwardly by peptide synthesis, provides an attractive alternative. In this study, TOAC was incorporated into a peptide derived from focal adhesion kinase (FAK), and the interaction of the peptide with the Src homology 3 (SH3) domain of Src kinase was studied, using paramagnetic NMR. Placing TOAC within the binding motif of the peptide has a considerable effect on the peptide-protein binding, lowering the affinity substantially. When the TOAC is positioned just outside the binding motif, the binding constant remains nearly unaffected. Although the SH3 domain binds weakly and transiently to proline-rich peptides from FAK, the interaction is not very dynamic and the relative position of the spin-label to the protein is well-defined. It is concluded that TOAC can be used to generate reliable paramagnetic NMR restraints. |
