title: claMP Tag: a versatile inline metal-binding platform based on the metal abstraction peptide.
abstract: Molecularly targeted research and diagnostic tools are essential to advancing understanding and detection of many diseases. Metals often impart the desired functionality to these tools, and conjugation of high-affinity chelators to proteins is carried out to enable targeted delivery of the metal. This approach has been much more effective with large lanthanide series metals than smaller transition metals. Because chemical conjugation requires additional processing and purification steps and yields a heterogeneous mixture of products, inline incorporation of a peptide tag capable of metal binding is a highly preferable alternative. Development of a transition metal binding tag would provide opportunity to greatly expand metal-based analyses. The metal abstraction peptide (MAP) sequence was genetically engineered into recombinant protein to generate the claMP Tag. The effects of this tag on recombinant epidermal growth factor (EGF) protein expression, disulfide bond formation, tertiary structural integrity, and transition metal incorporation using nickel were examined to confirm the viability of utilizing the MAP sequence to generate linker-less metal conjugates.
authors.affiliation: Department of Chemistry, The University of Kansas , Lawrence, Kansas 66045, United States.
authors.forename: Brittney J
authors.initials: BJ
authors.lastname: Mills
authors.forename: Qingxin
authors.initials: Q
authors.lastname: Mu
authors.forename: Mary E
authors.initials: ME
authors.lastname: Krause
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Laurence
mesh_terms: D001665:Binding Sites; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D066255:EGF Family of Proteins / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D009532:Nickel / Q000737:chemistry* / Q000494:pharmacology; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
chemical_list: D066255:EGF Family of Proteins; D010455:Peptides; D009532:Nickel