Now showing items 1-2 of 2

A bump-hole strategy for increased stringency of cell-specific metabolic labeling of rna

Nguyen, Kim; Kubota, Miles; Del Arco, Jon; Feng, Chao; Singha, Monika; Beasley, Samantha; Sakr, Jasmine; P. Gandhi, Sunil; Blurton-Jones, Mathew; Fernández Lucas, Jesus; C. Spitale, Robert (Corporación Universidad de la Costa, 2020-11-21)

Profiling RNA expression in a cell-specific manner continues to be a grand challenge in biochemical research. Bioorthogonal nucleosides can be utilized to track RNA expression; however, these methods currently have limitations ...
Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii

Acosta, Javier; Nguyen, Kim; C. Spitale, Robert; Fernández-Lucas, Jesús (Bioresource Technology, 2021)

Nowadays, enzymatic synthesis of nucleotides is an efficient and sustainable alternative to chemical methodologies. In this regard, after the biochemical characterization of wild-type and mutant uracil phosphoribosyltransferases ...

Envíos recientes

A bump-hole strategy for increased stringency of cell-specific metabolic labeling of rna
...
Nguyen, Kim | 2020-11-21

Profiling RNA expression in a cell-specific manner continues to be a grand challenge in biochemical research. Bioorthogonal nucleosides can be utilized to track RNA expression; however, these methods currently have limitations due to background and incorporation of analogs into undesired cells. Herein, we design and demonstrate that uracil phosphoribosyltransferase can be engineered to match 5-vinyluracil for cell-specific metabolic labeling of RNA with exceptional specificity and stringency.

LEER
Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii
...
Acosta, Javier | 2021

Nowadays, enzymatic synthesis of nucleotides is an efficient and sustainable alternative to chemical methodologies. In this regard, after the biochemical characterization of wild-type and mutant uracil phosphoribosyltransferases from Toxoplasma gondii (TgUPRT, TgUPRT2, and TgUPRT3), TgUPRT2 was selected as the optimal candidate (69.5 IU mg−1, UMP synthesis) for structure-guided immobilization onto Ni2+ chelate (MNiUPRT2) and onto glutaraldehyde-activated microparticles (MGlUPRT2). Among resulting derivatives, MNiUPRT23 (6127 IU g−1biocat; 92% retained activity; 3–5 fold enhanced stability at 50–60 °C) and MGlUPRT2N (3711 IU g−1biocat; 27% retained activity; 8–20 fold enhanced stability at 50–60 °C) displayed the best operability. Moreover, the enzymatic synthesis of different pyrimidine NMPs was performed. Finally, the reusability of both derivatives in 5-FUMP synthesis (MNiUPRT23, 80% retained activity after 7 cycles, 5 min; MGlUPRT2N, 70% retained activity after 10 cycles, 20 min) was carried out at short times. © 2021 Elsevier Ltd

LEER

Browsing by Author "C. Spitale, Robert"

A bump-hole strategy for increased stringency of cell-specific metabolic labeling of rna ﻿

Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii ﻿

Envíos recientes

A bump-hole strategy for increased stringency of cell-specific metabolic labeling of rna

Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii

A bump-hole strategy for increased stringency of cell-specific metabolic labeling of rna

Taylor-made production of pyrimidine nucleoside-5′-monophosphate analogues by highly stabilized mutant uracil phosphoribosyltransferase from Toxoplasma gondii