title: Site-selective labeling strategies for screening by NMR.
abstract: NMR based screening has become an important tool in the pharmaceutical industry. Methods that provide information on the location of small molecule binding sites on the surface of a drug target (e. g. SAR-by-NMR and related techniques) are of particular interest. In order to extend the applicability of such techniques to drug targets of higher molecular weight, selective labeling strategies may be employed. Dual-amino acid selective labeling and site directed non-native amino acid replacement (SNAAR) allow for the selective detection of NMR resonances of a specific amino acid residue. This results in significantly reduced spectral complexity, which not only enables application to higher molecular weight systems, but also eliminates the need for sequential resonance assignment in order to identify the binding site. Regio-selective (or segmental) labeling of an entire protein domain of a multi domain protein may also be achieved. Labeling only a selected part of a multi domain protein (e. g. a catalytic or ligand binding domain) is an attractive way to simplify the spectral interpretation without disturbing the system under study.
authors.affiliation: Biovitrum, Structural Chemistry, Stockholm, S-112 76, Sweden. johan.weigelt@biovitrum.com
authors.forename: Johan
authors.initials: J
authors.lastname: Weigelt
authors.forename: Mats
authors.initials: M
authors.lastname: Wikström
authors.forename: Johan
authors.initials: J
authors.lastname: Schultz
authors.forename: Maria J P
authors.initials: MJ
authors.lastname: van Dongen
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis* / Q000138:chemical synthesis; D001665:Binding Sites; D002247:Carbon Isotopes; D007553:Isotope Labeling / Q000379:methods*; D008024:Ligands; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D008024:Ligands; D009587:Nitrogen Isotopes