TY - JOUR AU - Sebőkné Nagy, Krisztina AU - Kóta, Zoltán AU - Kincses, András AU - Fazekas, Ákos Ferenc AU - Dér, András AU - László, Zsuzsanna AU - Páli, Tibor TI - Spin-Label Electron Paramagnetic Resonance Spectroscopy Reveals Effects of Wastewater Filter Membrane Coated with Titanium Dioxide Nanoparticles on Bovine Serum Albumin JF - MOLECULES J2 - MOLECULES VL - 28 PY - 2023 IS - 19 EP - 17 PG - 17 SN - 1420-3049 DO - 10.3390/molecules28196750 UR - https://m2.mtmt.hu/api/publication/34153921 ID - 34153921 N1 - Funding Agency and Grant Number: We would like to acknowledge the support provided by the National Research, Development, and Innovation Office of Hungary-PD-143268. [PD-143268]; National Research, Development, and Innovation Office of Hungary Funding text: We would like to acknowledge the support provided by the National Research, Development, and Innovation Office of Hungary-PD-143268. AB - The accumulation of proteins in filter membranes limits the efficiency of filtering technologies for cleaning wastewater. Efforts are ongoing to coat commercial filters with different materials (such as titanium dioxide, TiO2) to reduce the fouling of the membrane. Beyond monitoring the desired effect of the retention of biomolecules, it is necessary to understand what the biophysical changes are in water-soluble proteins caused by their interaction with the new coated filter membranes, an aspect that has received little attention so far. Using spin-label electron paramagnetic resonance (EPR), aided with native fluorescence spectroscopy and dynamic light scattering (DLS), here, we report the changes in the structure and dynamics of bovine serum albumin (BSA) exposed to TiO2 (P25) nanoparticles or passing through commercial polyvinylidene fluoride (PVDF) membranes coated with the same nanoparticles. We have found that the filtering process and prolonged exposure to TiO2 nanoparticles had significant effects on different regions of BSA, and denaturation of the protein was not observed, neither with the TiO2 nanoparticles nor when passing through the TiO2-coated filter membranes. LA - English DB - MTMT ER - TY - JOUR AU - Varga-Zsíros, Vanda AU - Migh, Ede AU - Marton, Annamária AU - Kóta, Zoltán AU - Vizler, Csaba AU - Tiszlavicz, László AU - Horváth, Péter AU - Török, Zsolt AU - Vigh, László AU - Balogh, Gábor AU - Péter, Mária TI - Development of a Laser Microdissection-Coupled Quantitative Shotgun Lipidomic Method to Uncover Spatial Heterogeneity JF - CELLS J2 - CELLS-BASEL VL - 12 PY - 2023 IS - 3 PG - 16 SN - 2073-4409 DO - 10.3390/cells12030428 UR - https://m2.mtmt.hu/api/publication/33607647 ID - 33607647 AB - Lipid metabolic disturbances are associated with several diseases, such as type 2 diabetes or malignancy. In the last two decades, high-performance mass spectrometry-based lipidomics has emerged as a valuable tool in various fields of biology. However, the evaluation of macroscopic tissue homogenates leaves often undiscovered the differences arising from micron-scale heterogeneity. Therefore, in this work, we developed a novel laser microdissection-coupled shotgun lipidomic platform, which combines quantitative and broad-range lipidome analysis with reasonable spatial resolution. The multistep approach involves the preparation of successive cryosections from tissue samples, cross-referencing of native and stained images, laser microdissection of regions of interest, in situ lipid extraction, and quantitative shotgun lipidomics. We used mouse liver and kidney as well as a 2D cell culture model to validate the novel workflow in terms of extraction efficiency, reproducibility, and linearity of quantification. We established that the limit of dissectible sample area corresponds to about ten cells while maintaining good lipidome coverage. We demonstrate the performance of the method in recognizing tissue heterogeneity on the example of a mouse hippocampus. By providing topological mapping of lipid metabolism, the novel platform might help to uncover region-specific lipidomic alterations in complex samples, including tumors. LA - English DB - MTMT ER - TY - JOUR AU - Réthi-Nagy, Zsuzsánna AU - Ábrahám, Edit AU - Udvardy, Katalin AU - Klement, Éva AU - Darula, Zsuzsanna AU - Pál, Margit AU - Katona, Robert L. AU - Tubak, Vilmos AU - Páli, Tibor AU - Kóta, Zoltán AU - Sinka, Rita AU - Udvardy, Andor AU - Lipinszki, Zoltán TI - STABILON, a Novel Sequence Motif That Enhances the Expression and Accumulation of Intracellular and Secreted Proteins JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 23 PY - 2022 IS - 15 PG - 18 SN - 1661-6596 DO - 10.3390/ijms23158168 UR - https://m2.mtmt.hu/api/publication/33063331 ID - 33063331 N1 - Funding Agency and Grant Number: Ministry of Human Capacities of Hungary [NTP-NFTO21-B-0221]; Aron Marton College; Forerunner Federation; Ministry of Finance [GINOP2.1.2-8-1-4-16-2017-00319]; National Research, Development and Innovation Office [K132155]; Hungarian Academy of Sciences [BO/00329/15, LP2017-7/2017]; EU Horizon 2020 SGA grant [739593] Funding text: This research was funded by the Ministry of Human Capacities of Hungary (NTP-NFTO21-B-0221), Aron Marton College and Forerunner Federation to Z.R.-N., Ministry of Finance (GINOP2.1.2-8-1-4-16-2017-00319) to V.T., National Research, Development and Innovation Office (K132155) to R.S. and Hungarian Academy of Sciences (Bolyai Fellowship (BO/00329/15) and Lendulet Program Grant (LP2017-7/2017)) to Z.L., Z.D. and E.K. have received support from the EU Horizon 2020 SGA (739593) grant. AB - The dynamic balance of transcriptional and translational regulation together with degron-controlled proteolysis shapes the ever-changing cellular proteome. While a large variety of degradation signals has been characterized, our knowledge of cis-acting protein motifs that can in vivo stabilize otherwise short-lived proteins is very limited. We have identified and characterized a conserved 13-mer protein segment derived from the p54/Rpn10 ubiquitin receptor subunit of the Drosophila 26S proteasome, which fulfills all the characteristics of a protein stabilization motif (STABILON). Attachment of STABILON to various intracellular as well as medically relevant secreted model proteins resulted in a significant increase in their cellular or extracellular concentration in mammalian cells. We demonstrate that STABILON acts as a universal and dual function motif that, on the one hand, increases the concentration of the corresponding mRNAs and, on the other hand, prevents the degradation of short-lived fusion proteins. Therefore, STABILON may lead to a breakthrough in biomedical recombinant protein production. LA - English DB - MTMT ER - TY - JOUR AU - Csoboz, Bálint AU - Gombos, Imre AU - Kóta, Zoltán AU - Dukic, Barbara AU - Klement, Éva AU - Varga-Zsíros, Vanda AU - Lipinszki, Zoltán AU - Páli, Tibor AU - Vigh, László AU - Török, Zsolt TI - The Small Heat Shock Protein, HSPB1, Interacts with and Modulates the Physical Structure of Membranes JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 23 PY - 2022 IS - 13 SN - 1661-6596 DO - 10.3390/ijms23137317 UR - https://m2.mtmt.hu/api/publication/32930192 ID - 32930192 AB - Small heat shock proteins (sHSPs) have been demonstrated to interact with lipids and modulate the physical state of membranes across species. Through these interactions, sHSPs contribute to the maintenance of membrane integrity. HSPB1 is a major sHSP in mammals, but its lipid interaction profile has so far been unexplored. In this study, we characterized the interaction between HSPB1 and phospholipids. HSPB1 not only associated with membranes via membrane-form-ing lipids, but also showed a strong affinity towards highly fluid membranes. It participated in the modulation of the physical properties of the interacting membranes by altering rotational and lat-eral lipid mobility. In addition, the in vivo expression of HSPB1 greatly affected the phase behavior of the plasma membrane under membrane fluidizing stress conditions. In light of our current find-ings, we propose a new function for HSPB1 as a membrane chaperone. © 2022 by the authors. Licensee MDPI, Basel, Switzerland. LA - English DB - MTMT ER - TY - JOUR AU - Veszelka, Szilvia AU - Mészáros, Mária AU - Porkoláb, Gergő AU - Szecskó, Anikó AU - Kondor, Nóra AU - Ferenc, Györgyi AU - Polgár, Tamás Ferenc AU - Katona, Gábor AU - Kóta, Zoltán AU - Kelemen, Lóránd AU - Páli, Tibor AU - Vigh, Judit Piroska AU - Walter, Fruzsina AU - Bolognin, Silvia AU - Schwamborn, Jens C. AU - Jan, Jeng-Shiung AU - Deli, Mária Anna TI - A Triple Combination of Targeting Ligands Increases the Penetration of Nanoparticles across a Blood-Brain Barrier Culture Model JF - PHARMACEUTICS J2 - PHARMACEUTICS VL - 14 PY - 2022 IS - 1 PG - 19 SN - 1999-4923 DO - 10.3390/pharmaceutics14010086 UR - https://m2.mtmt.hu/api/publication/32575627 ID - 32575627 LA - English DB - MTMT ER - TY - JOUR AU - Péter, Mária AU - Gudmann, Péter AU - Kóta, Zoltán AU - Török, Zsolt AU - Vigh, László AU - Glatz, Attila AU - Balogh, Gábor TI - Lipids and Trehalose Actively Cooperate in Heat Stress Management of Schizosaccharomyces pombe JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 22 PY - 2021 IS - 24 SN - 1661-6596 DO - 10.3390/ijms222413272 UR - https://m2.mtmt.hu/api/publication/32531214 ID - 32531214 N1 - Cited By :3 Export Date: 20 September 2022 LA - English DB - MTMT ER - TY - JOUR AU - Rehman, Ateeq Ur AU - Bashir, Faiza AU - Ayaydin, Ferhan AU - Kóta, Zoltán AU - Páli, Tibor AU - Vass, Imre TI - Proline is a quencher of singlet oxygen and superoxide both in in vitro systems and isolated thylakoids JF - PHYSIOLOGIA PLANTARUM J2 - PHYSIOL PLANTARUM VL - 172 PY - 2021 IS - 1 SP - 7 EP - 18 PG - 12 SN - 0031-9317 DO - 10.1111/ppl.13265 UR - https://m2.mtmt.hu/api/publication/31682225 ID - 31682225 AB - Proline is a versatile plant metabolite, which is produced in large amounts in plants exposed to osmotic and oxidative stress. Proline has been shown to provide protection against various reactive oxygen species (ROS), such as hydrogen peroxide and hydroxyl radicals. On the other hand, its protective effect against singlet oxygen has been debated, and it is considered ineffective against superoxide. Here we used various methods for the detection of singlet oxygen (electron paramagnetic resonance, EPR, spin trapping by 2,2,6,6-tetramethyl-4-piperidone, fluorescence probing by singlet oxygen sensor green, SOSG, and oxygen uptake due to chemical trapping) and superoxide (oxygen uptake due to oxygen reduction) in vitro and in isolated thylakoids. We demonstrated that proline does quench both singlet oxygen and superoxide in vitro. By comparing the effects of chemical scavengers and physical quenchers, we concluded that proline eliminates singlet oxygen via a physical mechanism, with a bimolecular quenching rate of ca. 1.5-4 10(6) M-1 s(-1). Our data also show that proline can eliminate superoxide in vitro in a process that is likely to proceed via an electron transfer reaction. We could also show that proline does quench both singlet oxygen and superoxide produced in isolated thylakoids. The scavenging efficiency of proline is relatively small on a molar basis, but considering its presence in high amounts in plant cells under stress conditions it may provide a physiologically relevant contribution to ROS scavenging, supplementing other nonenzymatic ROS scavengers of plant cells. LA - English DB - MTMT ER - TY - JOUR AU - Páli, Tibor AU - Kóta, Zoltán TI - Studying Lipid–Protein Interactions with Electron Paramagnetic Resonance Spectroscopy of Spin-Labeled Lipids JF - METHODS IN MOLECULAR BIOLOGY J2 - METHODS MOL BIOL VL - 2003 PY - 2019 SP - 529 EP - 561 PG - 33 SN - 1064-3745 DO - 10.1007/978-1-4939-9512-7_22 UR - https://m2.mtmt.hu/api/publication/30804384 ID - 30804384 N1 - Export Date: 17 September 2019 Correspondence Address: Páli, T.; Biological Research Centre, Institute of BiophysicsHungary; email: tpali@brc.hu Export Date: 24 September 2019 Correspondence Address: Páli, T.; Biological Research Centre, Institute of BiophysicsHungary; email: tpali@brc.hu Export Date: 25 September 2019 Correspondence Address: Páli, T.; Biological Research Centre, Institute of BiophysicsHungary; email: tpali@brc.hu Export Date: 27 September 2019 Correspondence Address: Páli, T.; Biological Research Centre, Institute of BiophysicsHungary; email: tpali@brc.hu Export Date: 12 February 2020 Correspondence Address: Páli, T.; Biological Research Centre, Institute of BiophysicsHungary; email: tpali@brc.hu Cited By :2 Export Date: 2 May 2021 Correspondence Address: Páli, T.; Biological Research Centre, Hungary; email: tpali@brc.hu AB - Spin label electron paramagnetic resonance (EPR) of lipid–protein interactions reveals crucial features of the structure and assembly of integral membrane proteins. Spin-label EPR spectroscopy is the technique of choice to characterize the protein solvating lipid shell in its highly dynamic nature, because the EPR spectra of lipids that are spin-labeled close to the terminal methyl end of their acyl chains display two spectral components, those corresponding to lipids directly contacting the protein and those corresponding to lipids in the bulk fluid bilayer regions of the membrane. In this chapter, typical spin label EPR procedures are presented that allow determination of the stoichiometry of interaction of spin-labeled lipids with the intramembranous region of membrane proteins or polypeptides, as well as the association constant of the spin-labeled lipid with respect to the host lipid. The lipids giving rise to a so-called immobile spectral component in the EPR spectrum of such samples are identified as the motionally restricted first-shell lipids solvating membrane proteins in biomembranes. Stoichiometry and selectivity are directly related to the structure of the intramembranous sections of membrane-associated proteins or polypeptides and can be used to study the state of assembly of such proteins in the membrane. Since these characteristics of lipid–protein interactions are discussed in detail in the literature (see ref. Marsh, Eur Biophys J 39:513–525, 2010 for a recent review), here we focus more on how to spin label model membranes and biomembranes and how to measure and analyze the two-component EPR spectra of spin-labeled lipids in phospholipid bilayers that contain proteins or polypeptides. After a description of how to prepare spin-labeled model and native biological membranes, we present the reader with computational procedures for determining the molar fraction of motionally restricted lipids when both, one or none of the pure isolated—mobile or immobile—spectral components are available. With these topics, this chapter complements a previous methodological paper (Marsh, Methods 46:83–96, 2008). The interpretation of the data is discussed briefly, as well as other relevant and recent spin label EPR techniques for studying lipid–protein interactions, not only from the point of view of lipid chain dynamics. © Springer Science+Business Media, LLC, part of Springer Nature 2019. LA - English DB - MTMT ER - TY - JOUR AU - Mészáros, Mária AU - Porkoláb, Gergő AU - Kiss, Lóránd AU - Pilbat, Ana Maria AU - Kóta, Zoltán AU - Kupihár, Zoltán AU - Kéri, Albert AU - Galbács, Gábor AU - Siklós, László AU - Tóth, András AU - Fülöp, Lívia AU - Czirjákné Csete, Mária AU - Sipos, Áron AU - Hulper, P AU - Sipos, Péter AU - Páli, Tibor AU - Rákhely, Gábor AU - Révész, Piroska AU - Deli, Mária Anna AU - Veszelka, Szilvia TI - Niosomes decorated with dual ligands targeting brain endothelial transporters increase cargo penetration across the blood-brain barrier JF - EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES J2 - EUR J PHARM SCI VL - 123 ET - 0 PY - 2018 SP - 228 EP - 240 PG - 13 SN - 0928-0987 DO - 10.1016/j.ejps.2018.07.042 UR - https://m2.mtmt.hu/api/publication/3399566 ID - 3399566 AB - Nanoparticles targeting transporters of the blood-brain barrier (BBB) are promising candidates to increase the brain penetration of biopharmacons. Solute carriers (SLC) are expressed at high levels in brain endothelial cells and show a specific pattern at the BBB. The aim of our study was to test glutathione and ligands of SLC transporters as single or dual BBB targeting molecules for nanovesicles. High mRNA expression levels for hexose and neutral amino acid transporting SLCs were found in isolated rat brain microvessels and our rat primary cell based co-culture BBB model. Niosomes were derivatized with glutathione and SLC ligands glucopyranose and alanine. Serum albumin complexed with Evans blue (67kDa), which has a very low BBB penetration, was selected as a cargo. The presence of targeting ligands on niosomes, especially dual labeling, increased the uptake of the cargo molecule in cultured brain endothelial cells. This cellular uptake was temperature dependent and could be decreased with a metabolic inhibitor and endocytosis blockers filipin and cytochalasin D. Making the negative surface charge of brain endothelial cells more positive with a cationic lipid or digesting the glycocalyx with neuraminidase elevated the uptake of the cargo after treatment with targeted nanocarriers. Treatment with niosomes increased plasma membrane fluidity, suggesting the fusion of nanovesicles with endothelial cell membranes. Targeting ligands elevated the permeability of the cargo across the BBB in the culture model and in mice, and dual-ligand decoration of niosomes was more effective than single ligand labeling. Our data indicate that dual labeling with ligands of multiple SLC transporters can potentially be exploited for BBB targeting of nanoparticles. LA - English DB - MTMT ER - TY - JOUR AU - Veszelka, Szilvia AU - Mészáros, Mária AU - Kiss, Lóránd AU - Kóta, Zoltán AU - Páli, Tibor AU - Hoyk, Zsófia AU - Bozsó, Zsolt AU - Fülöp, Lívia AU - Tóth, András AU - Rákhely, Gábor AU - Deli, Mária Anna TI - Biotin and glutathione targeting of solid nanoparticles to cross human brain endothelial cells JF - CURRENT PHARMACEUTICAL DESIGN J2 - CURR PHARM DESIGN VL - 23 PY - 2017 IS - 28 SP - 4198 EP - 4205 PG - 8 SN - 1381-6128 DO - 10.2174/1381612823666170727144450 UR - https://m2.mtmt.hu/api/publication/3251708 ID - 3251708 LA - English DB - MTMT ER -