Upregulation of an Artificial Zymogen by Proteolysis

Abstract

Regulation of enzymatic activity is vital to living organisms. Here, we report the development and the genetic optimization of an artificial zymogen requiring the action of a natural protease to upregulate its latent asymmetric transfer hydrogenase activity.

Functionalized Ultrasmall Fluorinated Graphene with High NIR Absorbance for Controlled Delivery of Mixed Anticancer Drugs

“…DOX is not only an effective anticancer drug, but also exhibits bright fluorescence (Figure S4), making it a good fluorescent dye to label FGO−CS to see whether the nanocarrier is taken into the cells. Flow cytometry is a powerful tool and we have employed it to accurately measure the labeled protein or nucleic acid in single cells . So the cellular uptake behavior of FGO−CS was first investigated by flow cytometry measurements of DOX fluorescence in Hela cells.…”

“…[25,26] Briefly,a bout 1.5 10 6 Hela cells per well were seeded and pre-incubated in DMEM medium (2 mL) at 37 8Cf or 24 hi na5% CO 2 humidified atmosphere. To prepare nano-size FGO co-modified with oxygen and fluorine, a newly developed oxidation strategy was employed to control the chemical composition and structure.…”

“…Cellular uptake investigation : Flow cytometry was used to determine the cellular uptake behavior of a sample, based on the method established in our previous work . Briefly, about 1.5×10 6 Hela cells per well were seeded and pre‐incubated in DMEM medium (2 mL) at 37 °C for 24 h in a 5 % CO 2 humidified atmosphere.…”

Exploring and Adapting the Molecular Selectivity of Artificial Metalloenzymes

“…For iridium-based ArMs, the majority of literature studies thus far have extensively focused on their use as artificial transfer hydrogenases (ATHase). [113][114][115][116][117][118][119][120][121][122][123][124][125][126][127][128][129][130][131][132] In these works, asymmetric transfer hydrogenation has been shown possible for a number of substrates that include imines, enones, ketones, and for NAD + reduction. One intriguing example of an ATHase that has been used with biological systems relies on a cell-surface display system that shuttles streptavidin into the E. coli periplasm ( Figure 5A).…”

Artificial imine reductases: developments and future directions

“…15 The thought behind this design is to combine the diverse reaction scope of synthetic organometallic catalysts with selectivity influenced by a secondary coordination sphere from the protein scaffold. 16 The incorporation of an organometallic complex inside a protein has several potential benefits including (i) compatibility with milder conditions, as for natural enzymes, 17,18 (ii) protection from inactivation by external factors such as thiols, water and oxygen, 19,20 (iii) increased rates of catalysis, 21,22 and (iv) improved enantioselectivity from the presence of an extensive and complex secondary coordination sphere that would otherwise be extremely challenging to achieve synthetically. 23,24 The organometallic catalysts used for these artificial imine reductases (artIREDs) tend to use formate in place of the more expensive nicotinamide cofactors as a source of hydride for the reduction.…”