| 摘要 |
The invention refers to a method for identifying at least one binder molecul e comprising the steps of: (a) choosing two amino acid types (AA1 and AA2) in a polypeptide or protein of interest, whereby AA2 at least once occurs directl y subsequent to AA1 in the amino acid sequence of the polypeptide or protein, defining an amino acid pair AA1-AA2; (b) labelling the two amino acid types (AA1 and AA2) in the polypeptide or protein of interest, whereby all AA1- residues is labelled with 13C and all AA2-residues with 15N; (c) generating a first HNCO-type NMR spectrum of the labelled polypeptide or protein from ste p (b), thereby identifying signals from the labelled amino acidpair AA1-AA2; ( d) contacting the labelled polypeptide or protein with a potential binder molecule or a mixture of binder molecules under conditions and sufficient ti me for allowing binding of the potential binder molecule(s) and the labelled polypeptide or protein; (e) generating a second HNCO-type NMR spectrum, or a 1H-15N correlation type NMR spectrum, of the mix from step (d), monitoring signals identified in step (c); (f) comparing the first and the second NMR spectra, whereby a chemical shift change of the signals identified in step ( c) between the two spectra indicates an interaction between the potential binde r moleculeand the labelled polypeptide or protein.
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