Mass Spectrometry Applications

Cunsolo, Vincenzo; Foti, Salvatore; Ner‐Kluza, Joanna; Drabik, Anna; Silberring, Jerzy; Muccilli, Vera; Saletti, Rosaria; Pawlak, Katarzyna; Harwood, Emma; Yu, Fang; Ciborowski, Paweł; Anczkiewicz, Robert; Altweg, Kathrin; Spoto, Giuseppe; Pawlaczyk, Aleksandra; Szynkowska, M. I.; Smoluch, Marek; Kwiatkowska, Dorota

doi:10.1002/9781119377368.ch8

Cited by 2 publications

(2 citation statements)

References 194 publications

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“…Because a detailed description of the capabilities of the different mass analyzers, hybrid instruments, and fragmentation methods is out of the purpose of the present chapter, the reader is referred to a recently published review [ 57 ].…”

Section: Ms-based Techniques For Protein Analysismentioning

confidence: 99%

VDACs Post-Translational Modifications Discovery by Mass Spectrometry: Impact on Their Hub Function

Pittalà

Nibali

Reina

et al. 2021

IJMS

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VDAC (voltage-dependent anion selective channel) proteins, also known as mitochondrial porins, are the most abundant proteins of the outer mitochondrial membrane (OMM), where they play a vital role in various cellular processes, in the regulation of metabolism, and in survival pathways. There is increasing consensus about their function as a cellular hub, connecting bioenergetics functions to the rest of the cell. The structural characterization of VDACs presents challenging issues due to their very high hydrophobicity, low solubility, the difficulty to separate them from other mitochondrial proteins of similar hydrophobicity and the practical impossibility to isolate each single isoform. Consequently, it is necessary to analyze them as components of a relatively complex mixture. Due to the experimental difficulties in their structural characterization, post-translational modifications (PTMs) of VDAC proteins represent a little explored field. Only in recent years, the increasing number of tools aimed at identifying and quantifying PTMs has allowed to increase our knowledge in this field and in the mechanisms that regulate functions and interactions of mitochondrial porins. In particular, the development of nano-reversed phase ultra-high performance liquid chromatography (nanoRP-UHPLC) and ultra-sensitive high-resolution mass spectrometry (HRMS) methods has played a key role in this field. The findings obtained on VDAC PTMs using such methodologies, which permitted an in-depth characterization of these very hydrophobic trans-membrane pore proteins, are summarized in this review.

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Section: Ms-based Techniques For Protein Analysismentioning

confidence: 99%

VDACs Post-Translational Modifications Discovery by Mass Spectrometry: Impact on Their Hub Function

Pittalà

Nibali

Reina

et al. 2021

IJMS

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“…Recent advancements in MS techniques, such as liquid chromatography-tandem MS (LC-MS/MS), have enabled more accurate and precise measurements of vitamin K in biological samples. 7,8 Nonetheless, further enhancements in sample preparation, ionization techniques, and data analysis methods are still necessary to overcome the challenges inherent in measuring vitamin K using MS.…”

Section: Introductionmentioning

confidence: 99%

Measurement of vitamin K and its derivatives in human plasma through enzyme-assisted liquid chromatography-tandem mass spectrometry assay

Demirel

Selek

2023

Eur J Mass Spectrom (Chichester)

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Vitamin K is an essential lipophilic vitamin that acts as a coenzyme in several metabolic pathways. Accurate measurement of apolar metabolites transported by lipoproteins in serum matrices requires high-recovery extractions of vitamin K and its derivatives following standardized protocols. Conventionally developed methods in this field have predominantly employed solid-phase extraction for the measurement of vitamin K and its derivatives. In this study, our objective was to develop an enzyme-assisted extraction method for the precise measurement of vitamin K and its derivatives. Our methodology involved mixing 450 µL of serum samples with 50 µL of an internal standard and 50 µL of a lipase enzyme solution. Following vortexing, the mixture was incubated at 37°C for 15 min to activate the enzymes. The enzyme reaction was subsequently quenched with a mixture of 250 µL of methanol and 1 mL of hexane, followed by centrifugation at 12,000 g for 5 min. The upper phase was collected, concentrated using a concentrator device, and dissolved in a 100 µL solution of methanol/acetone/isopropanol (7:1:1, v/v/v) for analysis. Spectrum analysis was performed using the open-source MZmine 3 software, and a reference interval was established using the Python programming language on the Google Colab platform. The developed method for measuring vitamin K and its derivatives exhibited limit of detection and limit of quantitation values of 0.005 and 0.01 ng/mL, respectively. In conclusion, our study presents a precise and reliable method for the measurement of vitamin K and its derivatives using enzyme-assisted extraction.

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Mass Spectrometry Applications

Cited by 2 publications

References 194 publications

VDACs Post-Translational Modifications Discovery by Mass Spectrometry: Impact on Their Hub Function

VDACs Post-Translational Modifications Discovery by Mass Spectrometry: Impact on Their Hub Function

Measurement of vitamin K and its derivatives in human plasma through enzyme-assisted liquid chromatography-tandem mass spectrometry assay

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