Quantification of Metabolites by NMR Spectroscopy in the Presence of Protein

Wallmeier, Jens; Samol, Claudia; Ellmann, Lisa; Zacharias, Helena U.; Vogl, Franziska C.; Garcia, Muriel; Dettmer, Katja; Oefner, Peter J.; Gronwald, Wolfram; Investigators, Gckd

doi:10.1021/acs.jproteome.7b00057

Cited by 25 publications

(31 citation statements)

References 38 publications

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“…We identified that the expression of AF1q was dramatically elevated in glomeruli from mice treated with ADR. The interaction between AF1q and Notch signaling was reported in the commitment intrathymic projenitros toward the T-cell lineage [25]. In recent years, the role of AF1q in neuronal cells has been confirmed by identification of AF1q spatiotemporal expression in brain and the ability of AF1q in transforming human embryonic kidney (HEK) cells into neuron-like cells [26, 27].…”

Section: Discussionmentioning

confidence: 99%

AF1q Contributes to Adriamycin-Induced Podocyte Injury by Activating Wnt/β-Catenin Signaling

Ren

Sun

et al. 2017

Kidney Blood Press Res

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Background/Aims: Injury of podocytes plays an important role in decline of glomerular filtration and proteinuria. It is well-known that proteinuria is associated with numerous chronic kidney diseases (CKD). However, the underlying mechanism of podocyte injury remains unclear. Methods: We used reverse transcription-quantitative PCR (RT-qPCR) to compare the expression level of the ALL1-fused from the chromosome 1q (AF1q) gene in mice and mouse podocytes (MPC5) with or without Adriamycin (ADR) treatment. The effects of AF1q on Wnt/ β-catenin signaling were investigated by determining the expressions of desmin, snail, WT1, nephrin and E-cadherin using western blotting. Results: We found that AF1q expression was elevated in podocytes treated with ADR than untreated cells. AF1q overexpression directly led to podocytes injury with increased levels of desmin and snail. Luciferase activity of TOPflash reporter was significantly increased in cells with AF1q overexpression than wild type cells whereas deletion of T-cell-factor-7 (TCF7) eliminated this effect. Immunoprecipitation assay evidenced that AF1q interacted with TCF7 and promoted both transcriptional and translational expressions of TCF7. Overexpression of AF1q increased protein expression of β-catenin. However, in podocytes with deletion of TCF7, AF1q was not able to promote β-catenin expression. Conclusion: Our findings demonstrated that aberrant expression of AF1q may activate Wnt/β-catenin signaling and result in podocyte injury.

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Section: Discussionmentioning

confidence: 99%

AF1q Contributes to Adriamycin-Induced Podocyte Injury by Activating Wnt/β-Catenin Signaling

Ren

Sun

et al. 2017

Kidney Blood Press Res

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“…Mostly, 1D NOESY pulse sequences with presaturation during relaxation and mixing time together with additional spoil gradients for water suppression are used. If biofluid specimens such as plasma or serum contain large amounts of macromolecules, they should be either ultra filtrated prior to measurement [6] or CPMG-type pulse sequences are recommended for the attenuation of macromolecular signals [7,8].…”

Section: Nmr Measurementsmentioning

confidence: 99%

Data Normalization in NMR-based Metabolomics

Zacharias¹,

Altenbuchinger²,

Gronwald³

2018

Preprint

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The aim of this article is to summarize recent bioinformatic and statistical developments applicable to NMR-based metabolomics. Extracting relevant information from large multivariate datasets by statistical data analysis strategies may be of considerable complexity. Typical tasks comprise for example classification of specimens, identification of differentially produced metabolites, and estimation of fold changes. In this context it is of prime importance to minimize contributions from unwanted biases and experimental variance prior to these analyses. This is the goal of data normalization. Therefore, special emphasize is given to different data normalization strategies. In the first part, we will discuss the requirements and the pros and cons for a variety of commonly applied strategies. In the second part, we will concentrate on possible solutions in case that the requirements for the standard strategies are not fulfilled. In the last part, very recent developments will be discussed that allow reliable estimation of metabolic signatures for sample classification without prior data normalization. In this contribution special emphasis will be given to techniques that have worked well in our hands.

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“…After 15 days of consecutive administration, half of the rat population was used to collect samples such as serum, kidney, and liver tissue, while the other half underwent an additional 15 days of recovery before the same samples were acquired. The samples were measured using 1 H-NMR CPMG (an experiment used to suppress signals from larger molecules, see below) [ 150 , 151 , 152 ] along with 1 H, 1 H-COSY, 1 H, 13 C-HSQC, and 1 H, 13 C-HMBC used for molecules assignment. The obtained spectra showed that the “RenqingMangjue” pill alters many metabolites, which are related to a variety of metabolic pathways including energy metabolism, amino acid metabolism, and lipid metabolism indicating potentially harmful effects on kidneys and liver.…”

Section: An Introduction To Nmr Spectroscopymentioning

confidence: 99%

NMR as a “Gold Standard” Method in Drug Design and Discovery

et al. 2020

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Studying disease models at the molecular level is vital for drug development in order to improve treatment and prevent a wide range of human pathologies. Microbial infections are still a major challenge because pathogens rapidly and continually evolve developing drug resistance. Cancer cells also change genetically, and current therapeutic techniques may be (or may become) ineffective in many cases. The pathology of many neurological diseases remains an enigma, and the exact etiology and underlying mechanisms are still largely unknown. Viral infections spread and develop much more quickly than does the corresponding research needed to prevent and combat these infections; the present and most relevant outbreak of SARS-CoV-2, which originated in Wuhan, China, illustrates the critical and immediate need to improve drug design and development techniques. Modern day drug discovery is a time-consuming, expensive process. Each new drug takes in excess of 10 years to develop and costs on average more than a billion US dollars. This demonstrates the need of a complete redesign or novel strategies. Nuclear Magnetic Resonance (NMR) has played a critical role in drug discovery ever since its introduction several decades ago. In just three decades, NMR has become a “gold standard” platform technology in medical and pharmacology studies. In this review, we present the major applications of NMR spectroscopy in medical drug discovery and development. The basic concepts, theories, and applications of the most commonly used NMR techniques are presented. We also summarize the advantages and limitations of the primary NMR methods in drug development.

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Quantification of Metabolites by NMR Spectroscopy in the Presence of Protein

Cited by 25 publications

References 38 publications

AF1q Contributes to Adriamycin-Induced Podocyte Injury by Activating Wnt/β-Catenin Signaling

AF1q Contributes to Adriamycin-Induced Podocyte Injury by Activating Wnt/β-Catenin Signaling

Data Normalization in NMR-based Metabolomics

NMR as a “Gold Standard” Method in Drug Design and Discovery

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