title: Applications of a peptide ligand for streptavidin: the Strep-tag.
abstract: The Strep-tag constitutes a nine amino acid-peptide that binds specifically to streptavidin and occupies the same pocket where biotin is normally complexed. Since the Strep-tag participates in a reversible interaction it can be applied for the efficient purification of corresponding fusion proteins on affinity columns with immobilized streptavidin. Elution of the bound recombinant protein can be effected under mild buffer conditions by competition with biotin or a suitable derivative. In addition, Strep-tag fusion proteins can be easily detected in immunochemical assays, like Western blots or ELISAs, by means of commercially available streptavidin-enzyme conjugates. The Strep-tag/streptavidin system has been systematically optimized over the past years, including the engineering of streptavidin itself. Structural insight into the molecular mimicry between the peptide and biotin was furthermore gained from X-ray crystallographic analysis. As a result the system provides a reliable and versatile tool in recombinant protein chemistry. Exemplary applications of the Strep-tag are discussed in this review.
authors.affiliation: Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Skerra
authors.forename: T G
authors.initials: TG
authors.lastname: Schmidt
mesh_terms: D000345:Affinity Labels / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D008024:Ligands; D046911:Macromolecular Substances; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D008024:Ligands; D046911:Macromolecular Substances; D009842:Oligopeptides; D011994:Recombinant Proteins; D019809:Streptavidin