TY - JOUR AU - Bruszel, Bella AU - Tóth-Molnár, Edit AU - Janáky, Tamás AU - Szabó, Zoltán TI - Sources of Variance in Human Tear Proteomic Samples: Statistical Evaluation, Quality Control, Normalization, and Biological Insight JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 25 PY - 2024 IS - 3 PG - 19 SN - 1661-6596 DO - 10.3390/ijms25031559 UR - https://m2.mtmt.hu/api/publication/34536265 ID - 34536265 AB - Human tear fluid contains numerous compounds, which are present in highly variable amounts owing to the dynamic and multipurpose functions of tears. A better understanding of the level and sources of variance is essential for determining the functions of the different tear components and the limitations of tear samples as a potential biomarker source. In this study, a quantitative proteomic method was used to analyze variations in the tear protein profiles of healthy volunteers. High day-to-day and inter-eye personal variances were observed in the tear volumes, protein content, and composition of the tear samples. Several normalization and outlier exclusion approaches were evaluated to decrease variances. Despite the intrapersonal variances, statistically significant differences and cluster analysis revealed that proteome profile and immunoglobulin composition of tear fluid present personal characteristics. Using correlation analysis, we could identify several correlating protein clusters, mainly related to the source of the proteins. Our study is the first attempt to achieve more insight into the biochemical background of human tears by statistical evaluation of the experimentally observed dynamic behavior of the tear proteome. As a pilot study for determination of personal protein profiles of the tear fluids of individual patients, it contributes to the application of this noninvasively collectible body fluid in personal medicine. LA - English DB - MTMT ER - TY - BOOK AU - Csősz, Éva AU - Darula, Zsuzsanna AU - Drahos, László AU - Emri, Miklós AU - Hunyadi-Gulyás, Éva AU - Janáky, Tamás AU - Juhász, Gábor AU - Kalló, Gergő AU - Kékesi, Adrienna Katalin AU - Klement, Éva AU - Márk, László AU - Medzihradszky, F. Katalin AU - Pettkó-Szandtner, Aladár AU - Rokobné Révész, Ágnes AU - Schlosser, Gitta (Vácziné) AU - Szabó, Zoltán AU - Tóth, Gábor AU - Turiák, Lilla TI - Bevezetés a proteomikába : a fehérjék korszerű vizsgálata PB - Semmelweis Egyetem CY - Budapest PY - 2023 SN - 9789633316009 UR - https://m2.mtmt.hu/api/publication/34407280 ID - 34407280 LA - Hungarian DB - MTMT ER - TY - JOUR AU - Harmati, Mária AU - Bukva, Mátyás AU - Dobra, Gabriella AU - Gyukity-Sebestyén, Edina AU - Böröczky, Timea AU - Szabó, Zoltán AU - Kónya, Zoltán AU - Horvath, Peter AU - Klekner, Almos AU - Buzás, Krisztina TI - Extracellular vesicle-mediated intercellular communication in cancer JF - EUROPEAN JOURNAL OF IMMUNOLOGY J2 - EUR J IMMUNOL VL - 53 PY - 2023 IS - S1 SP - 39 EP - 40 PG - 2 SN - 0014-2980 UR - https://m2.mtmt.hu/api/publication/34231255 ID - 34231255 N1 - Funding Agency and Grant Number: National Brain Research Program NAP 2.0; Szent-Gyorgyi Albert Research Fund (2021) by the University of Szeged; [GINOP-2.2.1-15-2017-00052]; [TKP2021-EGA09] Funding text: This study was supported by GINOP-2.2.1-15-2017-00052; "National Brain Research Program NAP 2.0"; Szent-Gyorgyi Albert Research Fund (2021) provided by the University of Szeged and TKP2021-EGA09. Supplement: 1 LA - English DB - MTMT ER - TY - JOUR AU - Senobar Tahaei, Seyyed Ashkan AU - Kulmány, Ágnes Erika AU - Minorics, Renáta AU - Kiss, Anita AU - Szabó, Zoltán AU - Germán, Péter AU - Szebeni, Gábor AU - Gémes, Nikolett AU - Mernyák, Erzsébet AU - Zupkó, István TI - Antiproliferative and Antimetastatic Properties of 16-Azidomethyl Substituted 3-O-Benzyl Estrone Analogs JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 24 PY - 2023 IS - 18 PG - 16 SN - 1661-6596 DO - 10.3390/ijms241813749 UR - https://m2.mtmt.hu/api/publication/34131836 ID - 34131836 N1 - Funding Agency and Grant Number: The authors thank Dora Bokor, PharmD, for proofreading the manuscript. Funding text: The authors thank Dora Bokor, PharmD, for proofreading the manuscript. AB - Four diastereomers of 16-azidomethyl substituted 3-O-benzyl estradiol (1–4) and their two estrone analogs (16AABE and 16BABE) were tested for their antiproliferative properties against human gynecological cancer cell lines. The estrones were selected for additional experiments based on their outstanding cell growth-inhibiting activities. Both compounds increased hypodiploid populations of breast cancer cells, and 16AABE elicited cell cycle disturbance as evidenced by flow cytometry. The two analogs substantially increased the rate of tubulin polymerization in vitro. 16AABE and 16BABE inhibited breast cancer cells’ migration and invasive ability, as evidenced by wound healing and Boyden chamber assays. Since both estrone analogs exerted remarkable estrogenic activities, as documented by a luciferase reporter gene assay, they can be considered as promising drug candidates for hormone-independent malignancies. LA - English DB - MTMT ER - TY - JOUR AU - Tököli, Attila AU - Bodnár, Brigitta AU - Bogár, Ferenc AU - Paragi, Gábor AU - Hetényi, Anasztázia AU - Bartus, Éva AU - Wéber, Edit AU - Hegedüs, Zsófia AU - Szabó, Zoltán AU - Kecskeméti, Gábor AU - Szakonyi, Gerda AU - Martinek, Tamás TI - Structural Adaptation of the Single-Stranded DNA-Binding Protein C-Terminal to DNA Metabolizing Partners Guides Inhibitor Design JF - PHARMACEUTICS J2 - PHARMACEUTICS VL - 15 PY - 2023 IS - 4 PG - 17 SN - 1999-4923 DO - 10.3390/pharmaceutics15041032 UR - https://m2.mtmt.hu/api/publication/33712712 ID - 33712712 N1 - Department of Medical Chemistry, University of Szeged, Szeged, H6720, Hungary ELKH-SZTE Biomimetic Systems Research Group, Eötvös Loránd Research Network (ELKH), Szeged, H6720, Hungary Institute of Physics, University of Pécs, Pécs, H7624, Hungary Department of Theoretical Physics, University of Szeged, Szeged, H6720, Hungary Institute of Pharmaceutical Analysis, University of Szeged, Szeged, H6720, Hungary Export Date: 8 September 2023 Correspondence Address: Martinek, T.A.; Department of Medical Chemistry, Hungary; email: martinek.tamas@med.u-szeged.hu AB - Single-stranded DNA-binding protein (SSB) is a bacterial interaction hub and an appealing target for antimicrobial therapy. Understanding the structural adaptation of the disordered SSB C-terminus (SSB-Ct) to DNA metabolizing enzymes (e.g., ExoI and RecO) is essential for designing high-affinity SSB mimetic inhibitors. Molecular dynamics simulations revealed the transient interactions of SSB-Ct with two hot spots on ExoI and RecO. The residual flexibility of the peptide–protein complexes allows adaptive molecular recognition. Scanning with non-canonical amino acids revealed that modifications at both termini of SSB-Ct could increase the affinity, supporting the two-hot-spot binding model. Combining unnatural amino acid substitutions on both segments of the peptide resulted in enthalpy-enhanced affinity, accompanied by enthalpy–entropy compensation, as determined by isothermal calorimetry. NMR data and molecular modeling confirmed the reduced flexibility of the improved affinity complexes. Our results highlight that the SSB-Ct mimetics bind to the DNA metabolizing targets through the hot spots, interacting with both of segments of the ligands. LA - English DB - MTMT ER - TY - JOUR AU - Dobra, Gabriella AU - Gyukity-Sebestyén, Edina AU - Bukva, Mátyás AU - Harmati, Mária AU - Nagy, Valentina AU - Szabó, Zoltán AU - Pankotai, Tibor AU - Klekner, Álmos AU - Buzás, Krisztina TI - MMP-9 as Prognostic Marker for Brain Tumours: A Comparative Study on Serum-Derived Small Extracellular Vesicles JF - CANCERS J2 - CANCERS VL - 15 PY - 2023 IS - 3 PG - 21 SN - 2072-6694 DO - 10.3390/cancers15030712 UR - https://m2.mtmt.hu/api/publication/33592853 ID - 33592853 N1 - Laboratory of Microscopic Image Analysis and Machine Learning, Institute of Biochemistry, Biological Research Centre, Eötvös Loránd Research Network (ELKH), Szeged, H-6726, Hungary Doctoral School of Interdisciplinary Medicine, Albert Szent-Györgyi Medical School, University of Szeged, Szeged, H-6720, Hungary Department of Immunology, Albert Szent-Györgyi Medical School, Faculty of Science and Informatics, University of Szeged, Szeged, H-6720, Hungary Department of Medical Chemistry, Albert Szent-Györgyi Medical School, University of Szeged, Szeged, H-6720, Hungary Institute of Pathology, Albert Szent-Györgyi Medical School, University of Szeged, Szeged, H-6720, Hungary Genome Integrity and DNA Repair Group, Hungarian Centre of Excellence for Molecular Medicine (HCEMM), University of Szeged, Szeged, H-6720, Hungary Department of Neurosurgery, Faculty of Medicine, University of Debrecen, Debrecen, H-4032, Hungary Export Date: 8 March 2023 Correspondence Address: Buzás, K.; Laboratory of Microscopic Image Analysis and Machine Learning, Hungary; email: buzas.krisztina@brc.hu AB - Matrix metalloproteinase-9 (MMP-9) degrades the extracellular matrix, contributes to tumour cell invasion and metastasis, and its elevated level in brain tumour tissues indicates poor prognosis. High-risk tissue biopsy can be replaced by liquid biopsy; however, the blood–brain barrier (BBB) prevents tumour-associated components from entering the peripheral blood, making the development of blood-based biomarkers challenging. Therefore, we examined the MMP-9 content of small extracellular vesicles (sEVs)—which can cross the BBB and are stable in body fluids—to characterise tumours with different invasion capacity. From four patient groups (glioblastoma multiforme, brain metastases of lung cancer, meningioma, and lumbar disc herniation as controls), 222 serum-derived sEV samples were evaluated. After isolating and characterising sEVs, their MMP-9 content was measured by ELISA and assessed statistically (correlation, paired t-test, Welch’s test, ANOVA, ROC). We found that the MMP-9 content of sEVs is independent of gender and age, but is affected by surgical intervention, treatment, and recurrence. We found a relation between low MMP-9 level in sEVs (<28 ppm) and improved survival (8-month advantage) of glioblastoma patients, and MMP-9 levels showed a positive correlation with aggressiveness. These findings suggest that vesicular MMP-9 level might be a useful prognostic marker for brain tumours. LA - English DB - MTMT ER - TY - JOUR AU - Kecskeméti, Gábor AU - Tóth-Molnár, Edit AU - Janáky, Tamás AU - Szabó, Zoltán TI - An Extensive Study of Phenol Red Thread as a Novel Non-Invasive Tear Sampling Technique for Proteomics Studies: Comparison with Two Commonly Used Methods JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 23 PY - 2022 IS - 15 PG - 20 SN - 1661-6596 DO - 10.3390/ijms23158647 UR - https://m2.mtmt.hu/api/publication/33046603 ID - 33046603 AB - Tear samples are considered in recent publications as easily, noninvasively collectible information sources for precision medicine. Their complex composition may aid the identification of biomarkers and the monitoring of the effectiveness of treatments for the eye and systemic diseases. Sample collection and processing are key steps in any analytical method, especially if subtle personal differences need to be detected. In this work, we evaluate the usability of a novel sample collection technique for human tear samples using phenol red threads (cotton thread treated with the pH indicator phenol red), which are efficiently used to measure tear volume in clinical diagnosis. The low invasiveness and low discomfort to the patients have already been demonstrated, but their applicability for proteomic sample collection has not yet been compared to other methods. We have shown, using various statistical approaches, the qualitative and quantitative differences in proteomic samples collected with this novel and two traditional methods using either glass capillaries or Schirmer’s paper strips. In all parameters studied, the phenol red threads proved to be equally or even more suitable than traditional methods. Based on detectability using different sampling methods, we have classified proteins in tear samples. LA - English DB - MTMT ER - TY - JOUR AU - Szabó, Márton Richárd AU - Pipicz, Márton AU - Sárközy, Márta AU - Bruszel, Bella AU - Szabó, Zoltán AU - Csont, Tamás Bálint TI - Diet-Induced Hypercholesterolemia Leads to Cardiac Dysfunction and Alterations in the Myocardial Proteome JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 23 PY - 2022 IS - 13 PG - 17 SN - 1661-6596 DO - 10.3390/ijms23137387 UR - https://m2.mtmt.hu/api/publication/32924828 ID - 32924828 N1 - Export Date: 18 October 2022 AB - Elevated blood cholesterol is a major risk factor for coronary heart disease. Moreover, direct effects on the myocardium also contribute to the adverse effects of hypercholesterolemia. Here, we investigated the effect of hypercholesterolemia on the cardiac proteome. Male Wistar rats were fed with a laboratory rodent chow supplemented with 2% cholesterol for 8 weeks to induce hypercholesterolemia. The protein expression data obtained from the proteomic characterization of left ventricular samples from normo- and hypercholesterolemic animals were subjected to gene ontology (GO) and protein interaction analyses. Elevated circulating cholesterol levels were accompanied by diastolic dysfunction in cholesterol-fed rats. The proteomic characterization of left ventricular samples revealed altered expression of 45 proteins due to hypercholesterolemia. Based on the Gene Ontology analysis, hypercholesterolemia was associated with disturbed expression of cytoskeletal and contractile proteins. Beta-actin was downregulated in the hypercholesterolemic myocardium, and established a prominent hub of the protein interaction network. Analysis of the unfiltered dataset revealed concordant downregulated expression patterns in proteins associated with the arrangement of the contractile system (e.g., cardiac-specific troponins and myosin complex), and in subunits of the mitochondrial respiratory chain. We conclude that the observed changes in the cardiac proteome may contribute to the development of diastolic dysfunction in hypercholesterolemia. LA - English DB - MTMT ER - TY - JOUR AU - Sáfár, Zsolt AU - Kecskeméti, Gábor AU - Molnár, Judit AU - Kurunczi, Anita AU - Szabó, Zoltán AU - Janáky, Tamás AU - Kis, Emese AU - Krajcsi, Péter TI - Inhibition of ABCG2/BCRP-mediated transport–correlation analysis of various expression systems and probe substrates JF - EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES J2 - EUR J PHARM SCI VL - 156 PY - 2021 PG - 11 SN - 0928-0987 DO - 10.1016/j.ejps.2020.105593 UR - https://m2.mtmt.hu/api/publication/31646463 ID - 31646463 N1 - Solvo Biotechnology, a Charles River Company, 52 Közép fasor, Szeged, H-6726, Hungary Department of Medical Chemistry, Faculty of Medicine, University of Szeged, Dóm tér 8, Szeged, H-6720, Hungary Solvo Biotechnology, a Charles River Company, 4-20 Irinyi J str, Budapest, H-1117, Hungary Faculty of Information Technology and Bionics, Pázmány Péter Catholic University, Práter str 50/a, Budapest, H-1083, Hungary Semmelweis University, Faculty of Health Sciences, Vas str 17, Budapest, H-1088, Hungary Cited By :3 Export Date: 12 January 2024 CODEN: EPSCE Correspondence Address: Krajcsi, P.; Solvo Biotechnology, 52 Közép fasor, Hungary; email: krajcsi@solvo.com AB - BCRP / ABCG2 is a key determinant of pharmacokinetics of substrate drugs. Several BCRP substrates and inhibitors are of low passive permeability, and the vesicular transport assay works well in this permeability space. Membranes were prepared from BCRP-HEK293, MCF-7/MX, and baculovirus-infected Sf9 cells with (BCRP-Sf9-HAM), and without (BCRP-Sf9) cholesterol loading. Km values for three substrates - estrone-3-sulfate, sulfasalazine, topotecan - correlated well between the four expression systems. In contrast, a 10-20-fold range in Vmax values was observed, with BCRP-HEK293 membranes possessing the largest dynamic range. IC50 values of the different test systems were similar to each other, with 94.4% of pairwise comparisons being within 3-fold. Substrate dependent inhibition showed somewhat greater variation, as 81.4% of IC50 values in the BCRP-HEK293 membranes were within 3-fold in pairwise comparisons. Overall, BCRP-HEK293 membranes demonstrated the highest activity. The IC50 values showed good concordance but substrate dependent inhibition was observed for some drugs. LA - English DB - MTMT ER - TY - JOUR AU - Costa, Roberta AU - Remigante, Alessia AU - Civello, Davide A. AU - Bernardinelli, Emanuele AU - Szabó, Zoltán AU - Morabito, Rossana AU - Marino, Angela AU - Sarikas, Antonio AU - Patsch, Wolfgang AU - Paulmichl, Markus AU - Janáky, Tamás AU - Miseta, Attila János AU - Nagy, Tamás AU - Dossena, Silvia TI - O-GlcNAcylation Suppresses the Ion Current IClswell by Preventing the Binding of the Protein ICln to α-Integrin JF - FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY J2 - FRONT CELL DEV BIOL VL - 8 PY - 2020 PG - 23 SN - 2296-634X DO - 10.3389/fcell.2020.607080 UR - https://m2.mtmt.hu/api/publication/31666189 ID - 31666189 N1 - * Megosztott szerzőség AB - O-GlcNAcylation is a post-translational modification of proteins that controls a variety of cellular processes, is chronically elevated in diabetes mellitus, and may contribute to the progression of diabetic complications, including diabetic nephropathy. Our previous work showed that increases in the O-GlcNAcylation of cellular proteins impair the homeostatic reaction of the regulatory volume decrease (RVD) after cell swelling by an unknown mechanism. The activation of the swelling-induced chloride current IClswell is a key step in RVD, and ICln, a ubiquitous protein involved in the activation of IClswell, is O-GlcNAcylated. Here, we show that experimentally increased O-GlcNAcylation of cellular proteins inhibited the endogenous as well as the ICln-induced IClswell current and prevented RVD in a human renal cell line, while decreases in O-GlcNAcylation augmented the current magnitude. In parallel, increases or decreases in O-GlcNAcylation, respectively, weakened or stabilized the binding of ICln to the intracellular domain of α-integrin, a process that is essential for the activation of IClswell. Mutation of the putative YinOYang site at Ser67 rendered the ICln-induced IClswell current unresponsive to O-GlcNAc variations, and the ICln interaction with α-integrin insensitive to O-GlcNAcylation. In addition, exposure of cells to a hypotonic solution reduced the O-GlcNAcylation of cellular proteins. Together, these findings show that O-GlcNAcylation affects RVD by influencing IClswell and further indicate that hypotonicity may activate IClswell by reducing the O-GlcNAcylation of ICln at Ser67, therefore permitting its binding to α-integrin. We propose that disturbances in the regulation of cellular volume may contribute to disease in settings of chronically elevated O-GlcNAcylation, including diabetic nephropathy. LA - English DB - MTMT ER -