Monitoring of successive phosphorylations of thymidine using free and immobilized human nucleoside/nucleotide kinases by Flow Injection Analysis with High-Resolution Mass Spectrometry

Ferey, Justine; Silva, David Da; Colas, Cyril; Nehmé, Reine; Lafite, Pierre; Roy, Vincent; Morin, Pascal; Daniellou, Richard; Agrofoglio, Luigi A.; Maunit, Benoı̂t

doi:10.1016/j.aca.2018.10.032

Cited by 6 publications

(5 citation statements)

References 44 publications

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“…The evaluation of K m with online reactions is simpler and preferable to off-line reactions. In the period under review, K m values were quantified online with CE-UV absorbance detection for a variety of enzymes including ecto-5-nucleotidase, human recombinant matrix metalloproteinase, human neutrophil elastase, bovine testicular hyaluronidase, histone deacetylase, thrombin, and human nucleoside/nucleotide kinases . In other reports, K m values were quantified online with CE-fluorescence detection for d -amino acid oxidase and BCR-ABL .…”

Section: Proteinsmentioning

confidence: 99%

“…In the period under review, K m values were quantified online with CE-UV absorbance detection for a variety of enzymes including ecto-5-nucleotidase, 54 human recombinant matrix metalloproteinase, 55 human neutrophil elastase, 55 bovine testicular hyaluronidase, 55 histone deacetylase, 56 thrombin, 52 and human nucleoside/nucleotide kinases. 57 In other reports, K m values were quantified online with CE-fluorescence detection for D-amino acid oxidase 58 and BCR-ABL. 59 Further, K m values have been determined using online CE-UV detection for immobilized enzymes like penicillinase, 47 acetylcholinesterase, 46,53 alanine aminotransferase, 49 trypsin, 44 and β-glucosidase.…”

Section: ■ Proteinsmentioning

confidence: 99%

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Challenging Bioanalyses with Capillary Electrophoresis

et al. 2019

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This report covers advances in capillary electrophoresis (CE) from January 2018 through September 2019. A summary of the literature during this time period is insightful. A search performed using the SciFinder Scholar® database for journal reports (limited to English) using the term capillary electrophoresis returned approximately 1,800 publications. Further analysis of this list, depicted in Figure 1, provided a snapshot of activity in biomolecular research. Classes of biomolecules most frequently associated with CE publications were proteins, drugs, DNA and metabolites. Another measure of the impact of CE is the translation of this technology into society. A search of patent activity illustrating this process of CE technology transfer returned 346 patents published in all languages, with a substantial contribution reported only in Chinese (198 patents) or English (98 patents). The versatility of CE for biological systems is exemplified by the rise of the technique in several areas. Metabolomics research involving measurements of large sets of molecules with subtle structural differences benefits from rapid separations achieved with high peak capacity and automated instruments. Single cell and sub-cellular analyses continue to progress in CE because of the size compatibility of the technique with the sample. Other examples of areas utilizing CE that are accelerating include portable and printable instrumentation, affinity interaction, as well as proteomics. As an established analytical tool, CE instrumentation and methods have been designed to be accessible and easily adopted by researchers with expertise in areas beyond the field of separations. Generally, publications including CE measurements either outline innovations in the technique or they are compelling applications of a mature analytical approach. The goal of this review, which is limited to 180 citations, is

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Section: Proteinsmentioning

confidence: 99%

Section: ■ Proteinsmentioning

confidence: 99%

Challenging Bioanalyses with Capillary Electrophoresis

et al. 2019

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“…However, in the case of immobilized enzymes, it is necessary to draw attention to parameters that are not associated with free enzyme catalysis: diffusion-related restrictions and decrease in enzyme mobility. These factors could affect the mobility of substrates and cofactors, the mass transfer of substrates and products [104]. However, the mass transfer resistance has been shown to decrease with increased flow rates and increased stirring.…”

Section: In Vitro Protein Phosphorylation By Immobilized Kinasesmentioning

confidence: 99%

Contemporary Enzyme-Based Methods for Recombinant Proteins In Vitro Phosphorylation

Slováková

Bı́lková

2021

Catalysts

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Phosphorylation is a reversible, enzyme-controlled posttranslational process affecting approximately one-third of all proteins in eukaryotic cells at any given time. Any deviation in the degree and/or site of phosphorylation leads to an abnormal conformation of proteins, resulting in a decline or loss of their function. Knowledge of phosphorylation-related pathways is essential for understanding the understanding of the disease pathogenesis and for the design of new therapeutic strategies. Recent availability of various kinases at an affordable price differs in activity, specificity, and stability and provides the opportunity of studying and modulating this reaction in vitro. We can exploit this knowledge for other applications. There is an enormous potential to produce fully decorated and active recombinant proteins, either for biomedical or cosmetic applications. Closely related is the possibility to exploit current achievements and develop new safe and efficacious vaccines, drugs, and immunomodulators. In this review, we outlined the current enzyme-based possibilities for in vitro phosphorylation of peptides and recombinant proteins and the added value that immobilized kinases provide.

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“…FIA-HRMS was successfully applied in identifying and detecting target and suspect psychoactive compounds in recreational drugs on the basis of accurate mass measurements . In addition, FIA-HRMS was employed in the quantitative analysis of multianalytes in tobacco and in enzyme kinetic studies to monitor the successive phosphorylation of thymidine. , …”

Section: Flow Injection Analysis-mass Spectrometry (Fia-ms)mentioning

confidence: 99%

“…33 In addition, FIA-HRMS was employed in the quantitative analysis of multianalytes in tobacco and in enzyme kinetic studies to monitor the successive phosphorylation of thymidine. 8,34 A major setback in the utilization of FIA-MS is matrix effects, and whether it is through ion enhancement or suppression, it compromises the analytical data. 35 Similar to LC-MS, using suitable internal standards, can correct for matrix effects to ensure that the analytical method is accurate and reliable.…”

Section: ■ Flow Injection Analysis-mass Spectrometry (Fia-ms)mentioning

confidence: 99%

Fast Quantification Without Conventional Chromatography, The Growing Power of Mass Spectrometry

et al. 2020

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Mass spectrometry (MS) in hyphenated techniques is widely accepted as the gold standard quantitative tool in life sciences. However, MS possesses intrinsic analytical capabilities that allow it to be a stand-alone quantitative technique, particularly with current technological advancements. MS has a great potential for simplifying quantitative analysis without the need for tedious chromatographic separation. Its selectivity relies on multistage MS analysis (MSn), including tandem mass spectrometry (MS/MS), as well as the ever-growing advancements of high-resolution MS instruments. This perspective describes various analytical platforms that utilize MS as a stand-alone quantitative technique, namely, flow injection analysis (FIA), matrix assisted laser desorption ionization (MALDI), including MALDI-MS imaging and ion mobility, particularly high-field asymmetric waveform ion mobility spectrometry (FAIMS). When MS alone is not capable of providing reliable quantitative data, instead of conventional liquid chromatography (LC)-MS, the use of a guard column (i.e., fast chromatography) may be sufficient for quantification. Although the omission of chromatographic separation simplifies the analytical process, extra procedures may be needed during sample preparation and clean-up to address the issue of matrix effects. The discussion of this manuscript focuses on key parameters underlying the uniqueness of each technique for its application in quantitative analysis without the need for a chromatographic separation. In addition, the potential for each analytical strategy and its challenges are discussed as well as improvements needed to render them as mainstream quantitative analytical tools. Overcoming the hurdles for fully validating a quantitative method will allow MS alone to eventually become an indispensable quantitative tool for clinical and toxicological studies.

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Monitoring of successive phosphorylations of thymidine using free and immobilized human nucleoside/nucleotide kinases by Flow Injection Analysis with High-Resolution Mass Spectrometry

Cited by 6 publications

References 44 publications

Challenging Bioanalyses with Capillary Electrophoresis

Challenging Bioanalyses with Capillary Electrophoresis

Contemporary Enzyme-Based Methods for Recombinant Proteins In Vitro Phosphorylation

Fast Quantification Without Conventional Chromatography, The Growing Power of Mass Spectrometry

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