title: A genetically encodable microtag for chemo-enzymatic derivatization and purification of recombinant proteins.
abstract: Efficient separation of recombinant polypeptides from proteins of the expression host and their subsequent derivatisation with functional chemical groups is essential for the success of many biological applications. Numerous tag systems have been developed to facilitate the purification procedure but only limited progress has been made in development of generic methods for targeted modification of proteins with functional groups. In this work, we present a novel 6 amino acid long C-terminal protein tag that can be selectively modified with functionalized derivatives of farnesyl isoprenoids by protein farnesyltransferase. The reaction could be performed in complex protein mixtures without detectable unspecific labeling. We demonstrate that this modification can be used to purify the target protein by over 800-fold in a single purification step using phase partitioning. Moreover, we show that the fluorescent group could be used to monitor the interaction of the derivatized proteins with other polypeptides.
authors.affiliation: Max-Planck-Institute for Molecular Physiology, Otto-Hahn-Str. 11, 44227 Dortmund, Germany.
authors.forename: Beatrice-Elena
authors.initials: BE
authors.lastname: Dursina
authors.forename: Reinhard
authors.initials: R
authors.lastname: Reents
authors.forename: Anca
authors.initials: A
authors.lastname: Niculae
authors.forename: Alexei
authors.initials: A
authors.lastname: Veligodsky
authors.forename: Reinhard
authors.initials: R
authors.lastname: Breitling
authors.forename: Konstantin
authors.initials: K
authors.lastname: Pyatkov
authors.forename: Herbert
authors.initials: H
authors.lastname: Waldmann
authors.forename: Roger S
authors.initials: RS
authors.lastname: Goody
authors.forename: Kirill
authors.initials: K
authors.lastname: Alexandrov
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D020559:Monomeric GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism; D017368:Protein Prenylation / Q000502:physiology*; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D013729:Terpenes / Q000138:chemical synthesis
chemical_list: D011994:Recombinant Proteins; D013729:Terpenes; D020559:Monomeric GTP-Binding Proteins