title: Cobalt(III)-Mediated Permanent and Stable Immobilization of Histidine-Tagged Proteins on NTA-Functionalized Surfaces.
abstract: We present the cobalt(III)-mediated interaction between polyhistidine (His)-tagged proteins and nitrilotriacetic acid (NTA)-modified surfaces as a general approach for a permanent, oriented, and specific protein immobilization. In this approach, we first form the well-established Co(2+) -mediated interaction between NTA and His-tagged proteins and subsequently oxidize the Co(2+) center in the complex to Co(3+) . Unlike conventionally used Ni(2+) - or Co(2+) -mediated immobilization, the resulting Co(3+) -mediated immobilization is resistant toward strong ligands, such as imidazole and ethylenediaminetetraacetic acid (EDTA), and washing off over time because of the high thermodynamic and kinetic stability of the Co(3+) complex. This immobilization method is compatible with a wide variety of surface coatings, including silane self-assembled monolayers (SAMs) on glass, thiol SAMs on gold surfaces, and supported lipid bilayers. Furthermore, once the cobalt center has been oxidized, it becomes inert toward reducing agents, specific and unspecific interactions, so that it can be used to orthogonally functionalize surfaces with multiple proteins. Overall, the large number of available His-tagged proteins and materials with NTA groups make the Co(3+) -mediated interaction an attractive and widely applicable platform for protein immobilization.
authors.affiliation: Department of New Materials and Biosystems, Max Planck Institute for Intelligent Systems, Heisenbergstrasse 3, 70569, Stuttgart, Germany. wegner@is.mpg.de.|Department of Biophysical Chemistry, University of Heidelberg, Im Neuenheimer Feld 253, 69120, Heidelberg, Germany. wegner@is.mpg.de.|Max Planck Institute for Polymer Research, Ackermannweg 10, 55128, Mainz, Germany. wegner@is.mpg.de.
authors.forename: Seraphine V
authors.initials: SV
authors.lastname: Wegner
authors.affiliation: Department of New Materials and Biosystems, Max Planck Institute for Intelligent Systems, Heisenbergstrasse 3, 70569, Stuttgart, Germany.|Department of Biophysical Chemistry, University of Heidelberg, Im Neuenheimer Feld 253, 69120, Heidelberg, Germany.
authors.forename: Franziska C
authors.initials: FC
authors.lastname: Schenk
authors.affiliation: Department of New Materials and Biosystems, Max Planck Institute for Intelligent Systems, Heisenbergstrasse 3, 70569, Stuttgart, Germany.|Department of Biophysical Chemistry, University of Heidelberg, Im Neuenheimer Feld 253, 69120, Heidelberg, Germany.
authors.forename: Joachim P
authors.initials: JP
authors.lastname: Spatz
mesh_terms: D003035:Cobalt / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry*; D007700:Kinetics; D009571:Nitrilotriacetic Acid / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D013499:Surface Properties
chemical_list: D055767:Immobilized Proteins; D011506:Proteins; C033223:polyhistidine; D003035:Cobalt; D006639:Histidine; D009571:Nitrilotriacetic Acid
keywords: biofunctionalization; cobalt; histidine; immobilization; surfaces and interfaces