WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection
Permanent URI for this collectionhttps://hdl.handle.net/11147/7150
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Conference Object Thermal Stability and Conformational Changes of Ubiquitin Upon Interaction with Few-Layer Graphene: An Optical Spectroscopy Approach(Springer, 2025) Gencay, Burcu; Guler, GunnurConference Object Reagent-Free Urea Determination From Hemodialysis Fluid: Development of FT-IR Spectroscopic Strategies(Springer, 2025) Akyuz, Ersed; Tanrisev, Mehmet; Guler, GunnurArticle Linking RNA Methylation to Structure: A Biophysical Perspective(Wiley, 2026) Akgul, Bunyamin; Guler, Gunnur; Saglam, Buket; Akkus, Onur; Akcaoz-Alasar, AzimeRecent epitranscriptomic studies show that ribonucleic acids (RNAs) are coated with an array of chemical modifications that dictate their cellular fate. Genetic, biochemical, and genomic approaches have been employed to elucidate the molecular details of RNA methylation, one of the most prevalent types of RNA modifications with significant implications for health and disease. Various biochemical approaches have been developed to identify RNA methylations both at the global and nucleotide resolution levels. However, simpler detection methods are needed to assess the global methylation status of synthetic or cellular RNAs. Although significant progress has been made in elucidating the factors involved in writing, erasing, or reading methylated epitopes or structures, the impact of these methyl moieties on the secondary structure of RNAs or macromolecular interactions remains to be fully understood. Typically, biophysical approaches, such as Fourier transformed-infrared (FT-IR) spectroscopy, circular dichroism (CD), and Raman spectroscopy, have been used to study the structures and interactions of macromolecules, including DNA and proteins. Although RNAs harbor similar chemical modifications or structure-mediated functions, the number of RNA studies that employ biophysical approaches is scarce. In this viewpoint article, we present a biophysical perspective that links RNA methylation to structure and propose that FT-IR analyses can be employed to examine global changes in the abundance of cellular RNA m(6)A marks. Additionally, we discuss the potential applications of biophysical approaches that may be employed to gain insight into methylation-mediated changes in RNA structures.Article Investigation of Few-Layer Graphene-Ubiquitin Interactions with Optical Spectroscopy Techniques(MDPI, 2025) Gencay, Burcu; Guler, GunnurUnderstanding the molecular mechanisms of protein-nanoparticle interactions is crucial for enabling the development of new applications in biomedicine and nanotechnology. Ubiquitin, an important and structurally small functional protein, plays a central role in numerous cellular processes. Therefore, in the current study, we focused on the few-layer graphene (FLG)-Ubiquitin complexes formed by exfoliating FLG structures using only water. Optical spectroscopic techniques (Raman, FT-IR, UV-Vis and circular dichroism) were employed to investigate these complexes on the molecular level. Overall, both CD and FT-IR data reveal that the formation of the FLG-Ubiquitin complexes occurred without inducing disordered structures in the protein. Based on the existence of a blue shift (hypsochromic shift) in the UV-Vis data, the presence of a single tyrosine and two phenylalanine residues in ubiquitin enables the detection of FLG-induced micro-environmental changes, particularly influencing the protein's beta-sheet and alpha-helix structures. The CD spectral results and CDPro quantitative estimations are in line with ATR FT-IR results, confirming the absence of disordered structure formation while altering the protein's chirality. UV-Vis and CD spectroscopy results revealed concentration-dependent trends consistent with FLG-protein interactions that preserve the overall protein structure. This study has potential applications in both academic research and practical usage, particularly in biomedicine and nanotechnology specifically for FLG.Article Citation - WoS: 1Citation - Scopus: 1Biophysical Assessment of Protein Stability in Ethanol-Stressed Environments via UV Absorption and Fluorescence Spectroscopies(Elsevier, 2026) Akyuz, Ersed; Vorob'ev, Mikhail M.; Guler, GunnurMaintaining the structure and functionality of proteins is crucial in applications ranging from food preservation to pharmaceutical formulation. Ethanol, while commonly used as a solvent and preservative, can induce structural changes in proteins depending on its concentration and the specific structure of the protein itself. This study investigates the structural effects of ethanol on three types of model proteins, namely bovine serum albumin (BSA), beta-Lactoglobulin (beta-Lg), and beta-Casein (beta-Cn), by using UV-Vis spectroscopy and fluorescence spectroscopy. The conformational responses of proteins in water-EtOH solutions of various ethanol concentrations (0-10 %, v/v) were analyzed through absorbance and emission spectral changes. At increasing ethanol concentration, UV absorption data showed distinct protein-dependent spectral changes. beta-Lg and beta-Cn exhibited strong hypochromism (an absorbance decrease of similar to 25 %) and red-shifting at 222 nm and 220 nm, respectively, indicating partial unfolding and solvent exposure of aromatic residues. BSA demonstrated subtle changes, and consistent quenching in fluorescence with a continuous blue-shifting to 330 nm, suggesting a moderate overall stability and local rearrangements in its structure. beta-Cn exhibited red-shifted fluorescence and quenching, reflecting its flexible, disordered structure and heterogeneous response to solvent conditions. Statistical analysis revealed that while fluorescence spectroscopy was highly sensitive to the intrinsic differences between proteins (p < 0.001), the ethanol-induced conformational changes were too subtle to be detected as a statistically significant treatment effect. The consistency of these trends indicates a rational underlying mechanism of interaction. This reflects the subtle nature of the effect at the tested concentrations rather than the absence of an effect. Moreover, these results unveil the protein-specific effects of ethanol and strongly emphasize the importance of solvent composition in maintaining protein integrity. Ethanol concentrations up to 5 % may offer protein stability whereas high ethanol levels (>= 5-10 %) promote structural perturbations. These results will be useful for both basic scientific research, such as biophysical studies and the advancement of optical techniques, and various industrial uses.Article Integrated Spectroscopic and Morphological Analyses Reveal Cellular Shifts in Gene-Silenced Melanoma CSCs(Nature Portfolio, 2025) Ozdil, Berrin; Guler, Gunnur; Ataman, Evren; Aktug, HuseyinIntratumoral heterogeneity remains a major barrier to durable cancer therapies, largely driven by the persistence of cancer stem cells (CSCs). In this study, we employed an integrated, multi-scale approach to investigate how melanoma CSCs respond to siRNA-mediated silencing of three key regulatory genes: KLF4, SHH, and HIF1 alpha. Using a combination of morphological, molecular, spectroscopic, and elemental analyses, we explored structural and biochemical consequences of gene knockdown. Gene silencing resulted in significant changes in cell shape and size, reduced F-actin organization, and decreased PFN1 expression, indicating a loss of stem-like properties. ATR-FTIR spectroscopy revealed shifts in biomolecular composition, notably a reduction in amide III intensity and an increase in lipid ester content. SEM-EDS point-based elemental analysis revealed SEM-EDS point-based elemental analysis revealed relative differences in carbon and nitrogen levels between selected central and peripheral regions of silenced and control cells, at the micron-scale working depth, reflecting broader elemental distribution trends rather than precise subcellular compartmentalization. XPS analysis further confirmed these differences, providing additional insights into the elemental composition of the cellular surface. The integration of FTIR spectroscopy into this study highlights the potential of infrared spectroscopy as a powerful tool in cancer research. These findings demonstrate that targeting critical regulatory pathways induces cytoskeletal and biochemical remodelling in melanoma CSCs, offering a multi-dimensional perspective on cellular plasticity.Article Citation - WoS: 2Citation - Scopus: 2Vibrational Spectroscopy Unveils Distinct Cell Cycle Features of Cancer Stem Cells in Melanoma(Nature Portfolio, 2025) Uslu, Bensu Ruya; Ozdil, Berrin; Tarhan, Enver; Ozcelik, Serdar; Aktug, Huseyin; Guler, GunnurCancer stem cells (CSCs) play a central role in melanoma growth, resistance to treatment, and relapse, however, their dynamic regulatory behavior remains poorly understood. Vibrational spectroscopy offers a unique, label-free approach to investigate cellular heterogeneity at the molecular level. Here, we explored the biochemical and regulatory dynamics of CSCs identified by using a time-course design, integrating infrared and Raman spectroscopies with cell cycle analysis and immunocytochemistry targeting the checkpoint proteins p16 and p21. CSCs, non-cancer stem cells (NCSCs), and bulk CHL-1 melanoma cells were monitored at 11, 24, 48, and 72 h. CSCs showed a steady S-phase with an early rise in p16 followed by a subsequent increase in p21 expression, indicating a dynamic state of cell cycle checkpoints. In contrast, NCSCs and CHL-1 cells showed more transient p16/p21 expression and CHL-1 exhibited a marked p16 increase at 24 h. Spectroscopic analysis revealed that CSCs exhibited distinct vibrational profiles, predominantly in the nucleic acid-, protein- and lipid-associated regions. These differences were further supported by principal component and hierarchical clustering analyses, which consistently distinguished CSCs from NCSCs. Our findings underline the potential of vibrational spectroscopy to sensitively detect CSC-specific regulatory patterns and support its use in detecting new therapeutic targets in melanoma.Article Citation - WoS: 2Citation - Scopus: 2Modulating Cancer Stem Cell Characteristics in CD133+ Melanoma Cells through Hif1α, KLF4, and SHH Silencing(Amer Chemical Soc, 2025) Ozdil, Berrin; Güler, Günnur; Avci, Cigir Biray; Calik-Kocaturk, Duygu; Gorgulu, Volkan; Uysal, Aysegul; Guler, Gunnur; Aktug, HuseyinMalignant melanoma is a highly aggressive form of skin cancer, partly driven by a subset of cancer stem cells (CSCs) with remarkable capacities for self-renewal, differentiation, and resistance to therapy. In this study, we examined how silencing three key genes-Hif1 alpha, KLF4, and SHH-affects CSC characteristics. Using small interfering RNA (siRNA)-based approaches, we observed significant changes at both the gene and protein levels, shedding light on how these pathways influence melanoma progression. Our results demonstrated that silencing these genes reduces the stem-like features of CSCs. Notably, Hif1 alpha silencing triggered a marked decrease in hypoxia-related gene expression, while targeting SHH led to a reduction in Gli1, a downstream effector of SHH signaling, highlighting its potential as a therapeutic target. We also observed changes in epigenetic markers such as HDAC9 and EP300, which play crucial roles in maintaining stemness and regulating gene expression. Interestingly, these interventions appeared to reprogram CSCs, pushing them toward a phenotype distinct from both traditional CSCs and non-stem cancer cells (NCSCs). Our findings emphasize the importance of targeting key signaling pathways in melanoma CSCs and underscore the value of mimicking the tumor microenvironment in experimental models. By revealing the dynamic plasticity of melanoma CSCs, this study offers fresh insights into potential therapeutic strategies, particularly using siRNA to modulate pathways associated with tumor progression and stem cell behavior.Article Citation - WoS: 2Citation - Scopus: 2Effect of Mirna Administration on Non-Small Cell Lung Cancer Cells Studied by Cellular Viability Assay and Atr-Ftir Spectroscopy Combined With Multivariate Data-Analysis(Elsevier, 2025) Dagdeviren, Melih; Guler, Gunnur; Guler, Egemen Erdem; Un, Cemal; Karabay-Yavasoglu, Nefise UlkuMicroRNAs (miRNAs), small non-coding RNAs, play a significant role in the regulation of gene expression by various mechanisms. Some miRNAs such as hsa-miR-145 (mir145), hsa-let-7a-1 (let7), hsa-miR-155 (mir155), and hsa-miR-29b (mir29b) are expressed at low levels in cancers and associated with proliferation, metastasis, invasion and apoptosis. In the current study, we aimed to investigate the effect of selected synthetic miRNAs and their combinations on the non-small cell lung cancer (NSCLC) cells (A549) by following the cell viability profile and alterations in the cellular biomolecules with biophysical features. After administration of commercial miRNAs and their various combinations to A549 cell line, each group was analyzed with cell viability assay and attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy combined with unsupervised multivariate analysis. Bioinformatics analysis was also performed to detect and to classify the target human genes obtained from the mirDB database. According to the cell viability results, the "mir29b + let7" combination and "mir155" significantly decreased the cancer cell viability whereas the "mir145 + mir29b" and "mir155 + mir145" combinations dramatically increased the cancer cell viability when compared to the control cells. The FTIR data revealed that administration of the "mir155", "mir29b + let7 + mir155", and "mir29b + let7" combinations caused a decrease in the contents of proteins, lipids and nucleic acids in A549 cells. This study suggests that those miRNA combinations might be potential targets for vaccines or miRNA-based therapies that can restore the miRNA activity and thus should be further evaluated to combat lung cancer with miRNA technology.