Methods in Molecular Biology

Héroux, Julie; Gharib, A.

doi:10.3109/9781420019957-3

Cited by 3 publications

(3 citation statements)

References 1 publication

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“…Figure 2 shows the results of in situ UV/Vis spectrophotometric assays to explore H 2 ‐driven FMN and FAD reduction by Hyd1 (produced and isolated in accord with the Supporting Information, Methods Section S1.2; reaction follows General Procedure A). The flavin moiety of FMN gives λ max at 445 nm and FAD at 450 nm, both of which bleach upon two‐electron reduction [29, 30] (Figure 2 A,B; see the Supporting Information, Figure S6 for spectra of fully reduced FMN). The decrease in oxidized flavin concentration over time was used to calculate initial enzyme activity (Figure 2 C,D).…”

Section: Methodsmentioning

confidence: 99%

E. coli Nickel‐Iron Hydrogenase 1 Catalyses Non‐native Reduction of Flavins: Demonstration for Alkene Hydrogenation by Old Yellow Enzyme Ene‐reductases**

et al. 2021

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A new activity for the [NiFe] uptake hydrogenase 1 of Escherichia coli (Hyd1) is presented. Direct reduction of biological flavin cofactors FMN and FAD is achieved using H 2 as a simple, completely atom-economical reductant. The robust nature of Hyd1 is exploited for flavin reduction across a broad range of temperatures (25-70 8C) and extended reaction times. The utility of this system as a simple, easy to implement FMNH 2 or FADH 2 regenerating system is then demonstrated by supplying reduced flavin to Old Yellow Enzyme "enereductases" to support asymmetric alkene reductions with up to 100 % conversion. Hyd1 turnover frequencies up to 20.4 min À1 and total turnover numbers up to 20 200 were recorded during flavin recycling. Scheme 1. Oxidized (left) and reduced (right) FMN or FAD cofactors.

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Section: Methodsmentioning

confidence: 99%

E. coli Nickel‐Iron Hydrogenase 1 Catalyses Non‐native Reduction of Flavins: Demonstration for Alkene Hydrogenation by Old Yellow Enzyme Ene‐reductases**

et al. 2021

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“…To further investigate the low crude purity observed when using MBHA resin but also TentaGel S RAM resin when the peptoid was synthesized using an automated set up, we inspected the effects of using different cleavage cocktails as an additional parameter. In the literature several protocols suggest cleaving conditions depending on the side chain functionality and the choice of resin when using solid-phase synthesis protocols (Bernatowicz et al, 1989;Sherrington, 1990;Walker, 2012;Novabiochem). Peptoids were cleaved under two different cleavage conditions, A (TFA:TIPS:H 2 O; 95:2.5:2.5) and B (TFA:DCM:anisole, 49:49:2) for 30 min.…”

Section: Cleavage Conditions Influence the Final Crude Purity Of Peptoidsmentioning

confidence: 99%

Synthesis of Peptoids Containing Multiple Nhtrp and Ntrp Residues: A Comparative Study of Resin, Cleavage Conditions and Submonomer Protection

et al. 2020

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Peptoids hold status as peptide-mimetics with versatile biological applications due to their proteolytic stability and structural diversity. Among those that have been studied in different biological systems, are peptoids with dominant balanced hydrophobic and charge distribution along the backbone. Tryptophan is an important amino acid found in many biologically active peptides. Tryptophan-like side chains in peptoids allow H-bonding, which is absent from the parent backbone, due to the unique indole ring. Furthermore, the rigid hydrophobic core and flat aromatic system influence the positioning in the hydrocarbon core and allows accommodating tryptophan-like side chains into the interfacial regions of bacterial membranes and causing bacterial membrane damage. Incorporating multiple tryptophan-like side chains in peptoids can be tricky and there is a lack of suitable, synthetic routes established. In this paper, we investigate the synthesis of peptoids rich in Nhtrp and Ntrp residues using different resins, cleavage conditions, and unprotected as well as tert-butyloxycarbonyl-protected amines suitable for automated solid-phase submonomer peptoid synthesis protocols.

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“…These changes can then cause the formation of new sequences, which finally results in the synthesis of novel amino acid structures. 18 Splicing occurs from the 5 0 to 3 0 side, and intronic base alterations of the 5 0 side typically result in defects in mRNA splicing and often result in the formation of defective mRNA molecules. Mutations that influence mRNA splicing usually contain 3 0 and 5 0 splicing locations.…”

mentioning

confidence: 99%

TP53 Intronic Mutations in Bovine Enzootic Hematuria–Associated Urinary Bladder Tumors

et al. 2012

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Tumor protein 53 (TP53) is a tumor suppressor gene that is frequently mutated in urinary bladder tumors in both humans and animals. In cattle, urinary bladder tumors have been reported as occurring spontaneously as well as in conjunction with bracken fern consumption-induced bovine enzootic hematuria (BEH). The goal of this study was to evaluate various types of bovine urinary bladder neoplasms for the presence of TP53 alterations, using the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method. DNA was extracted from both epithelial and mesenchymal urinary bladder tumor samples in cattle, associated with the chronic consumption of bracken fern. PCR was performed using primers targeted to exons 5 to 8, following electrophoresis and isolation, and the products were assessed by SSCP. Tumors in which alterations in the electrophoresis patterns were noted included hemangiomas, papillomas, and carcinomas in situ. Exemplars of these tumor types were selected for sequencing, and although no changes were noted in the 5 to 8 exon range, on either side of the designed primers for exon 6, there was some portion of intron 6 in which sequencing demonstrated a deletion of the thyamine nucleotide at position 9332. In summary, although mutations were not observed within exons 5 to 8, this represents the first report of an intronic mutation in the TP53 gene in association with bovine urinary bladder tumors. Mutations within introns can predispose tissues to the development of cancer, and therefore, a possible association between mutations of the introns of TP53 and the development of urinary bladder tumors in cattle with BEH should be further investigated.

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Methods in Molecular Biology

Cited by 3 publications

References 1 publication

E. coli Nickel‐Iron Hydrogenase 1 Catalyses Non‐native Reduction of Flavins: Demonstration for Alkene Hydrogenation by Old Yellow Enzyme Ene‐reductases**

E. coli Nickel‐Iron Hydrogenase 1 Catalyses Non‐native Reduction of Flavins: Demonstration for Alkene Hydrogenation by Old Yellow Enzyme Ene‐reductases**

Synthesis of Peptoids Containing Multiple Nhtrp and Ntrp Residues: A Comparative Study of Resin, Cleavage Conditions and Submonomer Protection

TP53 Intronic Mutations in Bovine Enzootic Hematuria–Associated Urinary Bladder Tumors

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