Cobalt(III) as a Stable and Inert Mediator Ion between NTA and His6‐Tagged Proteins

Wegner, Seraphine V.; Spatz, Joachim P.

doi:10.1002/anie.201210317

Cited by 89 publications

(91 citation statements)

References 33 publications

(50 reference statements)

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“…We recently described the Co 3+ ‐mediated interaction between NTA and His‐tagged proteins, which overcomes the above‐described limitations . The Co 3+ ‐mediated interaction between NTA and His‐tagged proteins is specific, thermodynamically stable, kinetically inert toward ligand exchange with chelators, and generally applicable (Scheme ).…”

Section: Introductionmentioning

confidence: 99%

“…Replacement of the divalent mediator ions with the Co 3+ ion has two important advantages: 1) the Co 3+ ion forms thermodynamically more stable complexes relative to the divalent counterparts (e.g., the dissociation constants for the NTA complex of Co 3+ , Co 2+ , Ni 2+ , and Zn 2+ ions are 10 −25 , 10 −11 , 10 −12 , and 10 −11 m respectively); 2) the Co 3+ ion in general forms kinetically inert complexes toward intersphere ligand exchange (e.g., although the hexa‐aqua complex of the Co 2+ ion has a ligand exchange rate of 3×10 6 s −1 , the corresponding Co 3+ complex has an exchange rate of less than 10 −6 s −1 ). The large increase in the thermodynamic and kinetic stabilities upon the oxidation of Co 2+ to Co 3+ has also been used for drug delivery, mechanistic studies in bioinorganic chemistry, the functionalization of porphyrin–phospholipid membranes with His‐tagged peptides, and protein labeling …”

Section: Introductionmentioning

confidence: 99%

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Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Wegner

Schenk

Spatz

2016

Chemistry A European J

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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.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Wegner

Schenk

Spatz

2016

Chemistry A European J

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“…This would allow enough space to accommodate two Co 2+ ions. This would be comparable to the coordination that is seen when His-tagged proteins bind to the metal of the nitriloacetic acid (NTA) affinity resin during protein purification [40]. The mechanism of the Co 2+ oxidation by H 2 O 2 most likely involves a peroxo complex at the dinuclear metal site.…”

Section: Discussionmentioning

confidence: 87%

A simple method to engineer a protein-derived redox cofactor for catalysis

Shin

Choi

Williamson

et al. 2014

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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The 6x-Histidine tag which is commonly used for purification of recombinant proteins was converted to a catalytic redox-active center by incorporation of Co2+. Two examples of the biological activity of this engineered protein-derived cofactor are presented. After inactivation of the natural diheme cofactor of MauG, it was shown that the Co2+-loaded 6xHis-tag could substitute for the hemes in the H2O2-driven catalysis of tryptophan tryptophylquinone biosynthesis. To further demonstrate that the Co2+-loaded 6xHis-tag could mediate long range electron transfer, it was shown that addition of H2O2 to the Co2+-loaded 6xHis-tagged Cu1+ amicyanin oxidizes the copper site which is 20 Å away. These results provide proof of principle for this simple method by which to introduce a catalytic redox-active site into proteins for potential applications in research and biotechnology.

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“…[17] In contrast to the more common nickel-mediated interaction, Co 3+ complexes between the Histagged protein and the NTAr esulted in ah igh thermodynamic and kinetic stability of the complex with extended longterm stability (t 1/2 % 7days). [17] In contrast to the more common nickel-mediated interaction, Co 3+ complexes between the Histagged protein and the NTAr esulted in ah igh thermodynamic and kinetic stability of the complex with extended longterm stability (t 1/2 % 7days).…”

Section: Key Principles and Recent Developmentsmentioning

confidence: 99%

Multivalent Chelators for In Vivo Protein Labeling

Wieneke

Tampé

2019

Angew Chem Int Ed

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With the advent of single‐molecule methods, chemoselective and site‐specific labeling of proteins evolved to become a central aspect in chemical biology as well as cell biology. Protein labeling demands high specificity, rapid as well as efficient conjugation, while maintaining low concentration and biocompatibility under physiological conditions. Generic methods that do not interfere with the function, dynamics, subcellular localization of proteins, and crosstalk with other factors are crucial to probe and image proteins in vitro and in living cells. Alternatives to enzyme‐based tags or autofluorescent proteins are short peptide‐based recognition tags. These tags provide high specificity, enhanced binding rates, bioorthogonality, and versatility. Here, we report on recent applications of multivalent chelator heads, recognizing oligohistidine‐tagged proteins. The striking features of this system has facilitated the analysis of protein complexes by single‐molecule approaches.

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Cobalt(III) as a Stable and Inert Mediator Ion between NTA and His6‐Tagged Proteins

Cited by 89 publications

References 33 publications

Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

Cobalt(III)‐Mediated Permanent and Stable Immobilization of Histidine‐Tagged Proteins on NTA‐Functionalized Surfaces

A simple method to engineer a protein-derived redox cofactor for catalysis

Multivalent Chelators for In Vivo Protein Labeling

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