Katalyse und Elektronentransfer in helikalen De‐novo‐Gerüststrukturen

Pinter, Tyler B. J.; Koebke, Karl J.; Pecoraro, Vincent L.

doi:10.1002/ange.201907502

Cited by 5 publications

(3 citation statements)

References 158 publications

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“…The ability to recapitulate the mechanistic requirements of enzymatic activity, therefore, is a thorough test of our understanding of these enzymes [6] . Our lab has a long‐standing history of investigating the metal binding properties of three‐fold symmetric metal‐binding sites within three‐stranded coiled coils (3SCC) [7] . These units are formed when sequences of 30 ( TRI ) or 37 ( Grand , GR ) amino acid long peptides containing heptad repeats assemble into parallel 3SCCs (Table 1).…”

Section: Peptide[a] Sequence Kdapp [M] Znii Cui Cuiimentioning

confidence: 99%

“…It is remarkable that this de novo designed protein can catalyze both the hydrolytic carbonic anhydrase activity using zinc as well as the redox conversion of nitrite to nitric oxide using copper, with no other modification than metal substitution. We have thus been exploring structural and functional models of the T2Cu center from CuNiR in our TRI scaffolds [7, 9] …”

Section: Peptide[a] Sequence Kdapp [M] Znii Cui Cuiimentioning

confidence: 99%

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Making or Breaking Metal‐Dependent Catalytic Activity: The Role of Stammers in Designed Three‐Stranded Coiled Coils

et al. 2020

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While many life-critical reactions would be infeasibly slow without metal cofactors, a detailed understanding of how protein structure can influence catalytic activity remains elusive. Using de novo designed three-stranded coiled coils (TRI and Grand peptides formed using a heptad repeat approach), we examine how the insertion of a three residue discontinuity, known as a stammer insert, directly adjacent to a (His) 3 metal binding site alters catalytic activity. The stammer, which locally alters the twist of the helix, significantly increases copper-catalyzed nitrite reductase activity (CuNiR). In contrast, the well-established zinc-catalyzed carbonic anhydrase activity (p-nitrophenyl acetate, pNPA) is effectively ablated. This study illustrates how the perturbation of the protein sequence using non-coordinating and non-acid base residues in the helical core can perturb metalloenzyme activity through the simple expedient of modifying the helical pitch adjacent to the catalytic center.

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Section: Peptide[a] Sequence Kdapp [M] Znii Cui Cuiimentioning

confidence: 99%

Section: Peptide[a] Sequence Kdapp [M] Znii Cui Cuiimentioning

confidence: 99%

Making or Breaking Metal‐Dependent Catalytic Activity: The Role of Stammers in Designed Three‐Stranded Coiled Coils

et al. 2020

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“…However, the ultimate goal in the artificial enzymes’ development remains their de novo design from scratch, to originate ad hoc catalysts able to promote the desired transformation [19–21] . For this approach, an adequate knowledge of the reaction mechanism and transition state is necessary, to identify an optimal protein backbone bearing amino acid residues arranged in a favorable position to accommodate the reaction partners [1,22] .…”

Section: Introductionmentioning

confidence: 99%

Design of Artificial Enzymes: Insights into Protein Scaffolds

2023

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The design of artificial enzymes has emerged as a promising tool for the generation of potent biocatalysts able to promote new-to-nature reactions with improved catalytic performances, providing a powerful platform for wide-ranging applications and a better understanding of protein functions and structures. The selection of an appropriate protein scaffold plays a key role in the design process. This review aims to give a general overview of the most common protein scaffolds that can be exploited for the generation of artificial enzymes. Several examples are discussed and categorized according to the strategy used for the design of the artificial biocatalyst, namely the functionalization of natural enzymes, the creation of a new catalytic site in a protein scaffold bearing a wide hydrophobic pocket and de novo protein design. The review is concluded by a comparison of these different methods and by our perspective on the topic.

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The pH‐Induced Selectivity Between Cysteine or Histidine Coordinated Heme in an Artificial α‐Helical Metalloprotein

et al. 2020

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De Novo metalloprotein design assesses the relationship between metal active site architecture and catalytic reactivity. Herein, we use an α‐helical scaffold to control the iron coordination geometry when a heme cofactor is allowed to bind to either histidine or cysteine ligands, within a single artificial protein. Consequently, we uncovered a reversible pH‐induced switch of the heme axial ligation within this simplified scaffold. Characterization of the specific heme coordination modes was done by using UV/Vis and Electron Paramagnetic Resonance spectroscopies. The penta‐ or hexa‐coordinate thiolate heme (9≤pH≤11) and the penta‐coordinate imidazole heme (6≤pH≤8.5) reproduces well the heme ligation in chloroperoxidases or cyt P450 monooxygenases and peroxidases, respectively. The stability of heme coordination upon ferric/ferrous redox cycling is a crucial property of the construct. At basic pHs, the thiolate mini‐heme protein can catalyze O2 reduction when adsorbed onto a pyrolytic graphite electrode.

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Katalyse und Elektronentransfer in helikalen De‐novo‐Gerüststrukturen

Cited by 5 publications

References 158 publications

Making or Breaking Metal‐Dependent Catalytic Activity: The Role of Stammers in Designed Three‐Stranded Coiled Coils

Making or Breaking Metal‐Dependent Catalytic Activity: The Role of Stammers in Designed Three‐Stranded Coiled Coils

Design of Artificial Enzymes: Insights into Protein Scaffolds

The pH‐Induced Selectivity Between Cysteine or Histidine Coordinated Heme in an Artificial α‐Helical Metalloprotein

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