Engineering Spatial Orthogonality into Protein Translation

Fossen, Elise M. Van; Bednar, Riley M.; Mehl, Ryan A.

doi:10.1021/acs.biochem.9b00569

Cited by 2 publications

(1 citation statement)

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“…Compartmentalization confines reactions to specific subcellular compartments or organelles, helping avoid crossreactions (Pei et al, 2022). Spatial segregation is achieved through protein-protein interactions or through the organization of enzymes in macromolecule complexes (Van Fossen et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

EngineeringCandida boidiniiformate dehydrogenase for activity with NMN(H)

Vainstein,

Banta

2024

Preprint

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Multi-step enzymatic reaction cascades often involve cofactors that serve as electron donors/acceptors in addition to the primary substrates. The co-localization of cascades can lead to cross-talk and competition, which can be unfavorable for the production of a targeted product. Orthogonal pathways allow reactions of interest to operate independently from the metabolic reactions within a cell; non-canonical cofactor analogs have been explored as a means to create these orthogonal pathways. Here, we aimed to engineer the formate dehydrogenase from Candid boidinii (CbFDH) for activity with the non-canonical cofactor nicotinamide adenine mononucleotide (NMN(H)). We used PyRosetta and structural alignment to design mutations that enable CbFDH to use NMN+ for the oxidation of formate. Although the suggested mutations did not result in enhanced activity with NMN+, we found that PyRosetta was able to easily design single mutations that disrupted all enzymatic activity.

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Section: Introductionmentioning

confidence: 99%