Martina Marchetti‐Deschmann

University of Vienna

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Evaluation of Pseudotrypsin Cleavage Specificity Towards Proteins by MALDI-TOF Mass Spectrometry

Protein and Peptide Letters (2015)

Filip Dyčka Vojtěch Franc Petr Fryčák Martin Raus Pavel Řehulka

Trypsin is a protease, which is commonly used for the digestion of protein samples in proteomic experiments. The process of trypsin autolysis is known to produce autolytic peptides as well as active enzyme forms with one or more intra-chain splits. In consequence, their variable presence can influence the digestion of a protein substrate in the reaction mixture. Besides two major and well-studied forms named β-trypsin and α-trypsin, there are also other active trypsin forms known such as γ-trypsin and pseudotrypsin (ψ-trypsin). In this work, the cleavage specificity of ψ-trypsin was evaluated using in-gel digestion of protein standards followed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and tandem mass spectrometry (MS/MS) analyses of the resulting peptides. The numbers of produced and matching peptides were similar to those obtained using α-/β-trypsin. The same experience was obtained with a real complex protein sample from rat urine. In previous reports, ψ-trypsin was supposed to generate non-specific cleavages, which has now been reevaluated. Purified ψ-trypsin cleaved all analyzed proteins preferentially on the C-terminal side of Lys and Arg residues in accordance with the canonical tryptic cleavage. However, a minor nonspecific cleavage performance was also registered (particularly after Tyr and Phe), which was considerably higher than in the case of trypsin itself.

Cleavage (geology)

MALDI imaging

10.2174/0929866522666151008151617

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Citations (9)

nES-DMA with Charge-reduction based on Soft X-ray Radiation: Analysis of a Recombinant Monoclonal Antibody

Journal of Chromatography B (2021)

Nicole Engel Nicole Puffler Martina Marchetti‐Deschmann Günter Allmaier Victor U. Weiss

Due to the fast growing importance of monoclonal antibodies in biomedical research, bioanalytics and human therapy, sensitive, fast and reliable methods are needed to monitor their production, target their characteristics, and for their final quality control. Application of a nano electrospray (nES) with soft X-ray radiation (SXR) based charge reduction and differential mobility analysis (DMA, aka nano electrospray gas-phase electrophoretic mobility molecular analysis, nES GEMMA) allows the size-separation and detection of macromolecules and (bio-)nanoparticles from a few nm up to several hundreds of nm in diameter in a native-like environment. The current study focuses on the analysis of a 148 kDa recombinant monoclonal antibody (rmAb) with the above mentioned instrumental setup and applying an universal detector, i.e. a water-based condensation particle detector (CPC). Next to the intact rmAb, its aggregates and fragment products after digestion with IdeS protease were analyzed. Additionally, influence of temperature treatment and pH variation on the stability of the rmAb was monitored. In this context, changes in electrophoretic mobility diameter (EMD) values, peak shape, and signal intensity based on particle numbers were of interest. Molecular weights calculated by application of a correlation derived from respective standard protein compounds were compared to mass spectrometric values and were found to be in good accordance. To conclude, we demonstrate that nES-DMA is a valuable tool in the characterization and quality control of rmABs.

10.1016/j.jchromb.2021.122925

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Moving towards green lubrication: tribological behavior and chemical characterization of spent coffee grounds oil

Green Chemistry Letters and Reviews (2023)

Jessica Pichler Rosa Maria Eder Lukas Widder Markus Varga Martina Marchetti‐Deschmann

With the EU aiming for net-zero greenhouse gas emissions by 2050, conventional production cycles must be transformed into cradle-to-cradle approaches. Spent coffee grounds are often dumped in landfills, with their potential as high-quality feedstock for biofuel or bio-lubricant production. Spent coffee grounds oil (SCGO) was investigated for its physicochemical properties while having more free acid groups compared to the reference polyalphaolefin 8 (PAO 8), which may cause faster oxidation. TGA results displayed comparable thermal stability of SCGO and PAO 8 for inert/oxidative atmosphere. The oil composition was characterized by ATR-FTIR, elemental analysis, and GC-EI-MS, where a higher oxygen content was found for SCGO, referring to functional ester/acid groups. The tribological behavior of SCGO was studied as lubricant base oil and as a 5% additive in PAO 8. The condition of fresh and tribologically used oils was investigated with High-Resolution-ESI-MS, and the worn surfaces were evaluated by light microscopy and topographic analysis. The results showed a superior friction coefficient of pure SCGO (µ = 0.092) to PAO 8 (µ = 0.129). The 5% SCGO additive in PAO 8 (µ = 0.095) could significantly reduce friction compared to pure PAO 8 on an unpolished 100Cr6 surface.

Inert gas

Thermal Stability

10.1080/17518253.2023.2215243

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Development of a defined medium for the heterotrophic cultivation of Metallosphaera sedula

Extremophiles (2024)

Viktor Laurin Sedlmayr Maximilian Luger Ernst Pittenauer Martina Marchetti‐Deschmann Laura Kronlachner

Abstract The heterotrophic cultivation of extremophilic archaea still heavily relies on complex media. However, complex media are associated with unknown composition, high batch-to-batch variability, potential inhibiting and interfering components, as well as regulatory challenges, hampering advancements of extremophilic archaea in genetic engineering and bioprocessing. For Metallosphaera sedula , a widely studied organism for biomining and bioremediation and a potential production host for archaeal ether lipids, efforts to find defined cultivation conditions have still been unsuccessful. This study describes the development of a novel chemically defined growth medium for M. sedula . Initial experiments with commonly used complex casein-derived media sources deciphered Casamino Acids as the most suitable foundation for further development. The imitation of the amino acid composition of Casamino Acids in basal Brock medium delivered the first chemically defined medium. We could further simplify the medium to 5 amino acids based on the respective specific substrate uptake rates. This first defined cultivation medium for M. sedula allows advanced genetic engineering and more controlled bioprocess development approaches for this highly interesting archaeon.

Bioplastic

10.1007/s00792-024-01348-0

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Visualizing fungal metabolites during mycoparasitic interaction by MALDI mass spectrometry imaging

PROTEOMICS (2016)

Matthias Holzlechner Sonja Reitschmidt Sabine Gruber Susanne Zeilinger Martina Marchetti‐Deschmann

Studying microbial interactions by MALDI mass spectrometry imaging (MSI) directly from growing media is a difficult task if high sensitivity is demanded. We present a quick and robust sample preparation strategy for growing fungi (Trichoderma atroviride, Rhizoctonia solani) on glass slides to establish a miniaturized confrontation assay. By this we were able to visualize metabolite distributions by MALDI MSI after matrix deposition with a home-built sublimation device and thorough recrystallization. We present for the first time MALDI MSI data for secondary metabolite release during active mycoparasitism.

Mass spectrometry imaging

10.1002/pmic.201500510

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Citations (34)

Critical illness-related corticosteroid insufficiency (CIRCI) in dogs with systemic inflammatory response syndrome (SIRS)

Journal of Veterinary Internal Medicine (2019)

Alessio Pierini Martina Marchetti‐Deschmann G. Favilla Eleonora Gori Ilaria Lippi

Inflammatory response

Source

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Comparison of two downstream processes for efficient and sustainable antivenom preparation in terms of yield, final product purity and virus-inactivating potential

Tihana Kurtović Marija Brgles Maja Lang Balija Dora Sviben Stephanie Steinberger

Antivenom

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Phosphonate coating of SiO2 nanoparticles abrogates inflammatory effects and local changes of the lipid composition in the rat lung: a complementary bioimaging study

Particle and Fibre Toxicology (2018)

Mandy Großgarten Matthias Holzlechner Antje Vennemann Anna Balbekova Karin Wieland

The well-known inflammatory and fibrogenic changes of the lung upon crystalline silica are accompanied by early changes of the phospholipid composition (PLC) as detected in broncho-alveolar lavage fluid (BALF). Amorphous silica nanoparticles (NPs) evoke transient lung inflammation, but their effect on PLC is unknown. Here, we compared effects of unmodified and phosphonated amorphous silica NP and describe, for the first time, local changes of the PLC with innovative bioimaging tools. Unmodified (SiO2-n), 3-(trihydroxysilyl) propyl methylphosphonate coated SiO2-n (SiO2-p) as well as a fluorescent surrogate of SiO2-n (SiO2-FITC) nanoparticles were used in this study. In vitro toxicity was tested with NR8383 alveolar macrophages. Rats were intratracheally instilled with SiO2-n, SiO2-p, or SiO2-FITC, and effects on lungs were analyzed after 3 days. BALF from the right lung was analyzed for inflammatory markers. Cryo-sections of the left lung were subjected to fluorescence microscopy and PLC analyses by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MS), Fourier transform infrared microspectroscopy (FT-IR), and tandem mass spectrometry (MS/MS) experiments. Compared to SiO2-p, SiO2-n NPs were more cytotoxic to macrophages in vitro and more inflammatory in the rat lung, as reflected by increased concentration of neutrophils and protein in BALF. Fluorescence microscopy revealed a typical patchy distribution of SiO2-FITC located within the lung parenchyma and alveolar macrophages. Superimposable to this particle distribution, SiO2-FITC elicited local increases of phosphatidylglycerol (PG) and phosphatidylinositol (PI), whereas phoshatidylserine (PS) and signals from triacylgyceride (TAG) were decreased in the same areas. No such changes were found in lungs treated with SiO2-p or particle-free instillation fluid. Phosphonate coating mitigates effects of silica NP in the lung and abolishes their locally induced changes in PLC pattern. Bioimaging methods based on MALDI-MS may become a useful tool to investigate the mode of action of NPs in tissues.

Parenchyma

Nanotoxicology

Silicon dioxide

10.1186/s12989-018-0267-z

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Citations (18)

Epigenetic Control of Mitochondrial Fission Enables Self-Renewal of Stem-like Tumor Cells in Human Prostate Cancer

Cell Metabolism (2019)

Gianluca Civenni Roberto Bosotti Andrea Timpanaro Ramiro Vázquez Jessica Merulla

10.1016/j.cmet.2019.05.004

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Citations (84)

ChemInform Abstract: Synthesis of New Tricyclic Spirobispidines via Ring Closing Metathesis Reaction.

ChemInform (2008)

Christian Hametner D. Dangl K. Mereiter Martina Marchetti‐Deschmann Johannes M. Froehlich

Abstract ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

Ring-Closing Metathesis

Tricyclic

Salt metathesis reaction

10.1002/chin.200812174

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