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    Bimolecular fluorescence complementation assay to explore protein- protein interactions of the Yersinia virulence factor YopM
    (Academic Press Inc Elsevier Science, 2021) Ugurlu, Ozge; Evran, Serap
    Yersinia outer protein M (YopM) is one of the effector proteins and essential for virulence. YopM is delivered by the Yersinia type III secretion system (T3SS) into the host cell, where it shows immunosuppressive effect through interaction with host proteins. Therefore, protein-protein interactions of YopM is significant to understand its molecular mechanism. In this study, we aimed to explore protein protein interactions of YopM with the two components of T3SS, namely LcrV and LcrG. We used bimolecular fluorescence complementation (BiFC) assay and monitored the reassembly of green fluorescence protein in Escherichia coli. As an indicator of the protein-protein interaction, we monitored the in vivo reconstitution of fluorescence by measuring fluorescence intensity and imaging the cells under fluorescence microscope. We showed, for the first time, that YopM interacts with LcrG, but not with LcrV. Here, we propose BiFC assay as a simple method to screen novel interaction partners of YopM. (c) 2021 Elsevier Inc. All rights reserved.
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    Nucleic Acid and Peptide Aptamers as Potential Antiviral Drugs
    (Bentham Science Publishers, 2021) Evran, Serap; Uğurlu, Özge; Man, Ezgi; Gültan, Merve; Özyurt, Canan
    Aptamers with target-specific binding properties have emerged as an alternative to antibodies. Nucleic acid aptamers are short single-stranded oligonucleotides that can fold into unique three-dimensional structures. Nucleic acid aptamers are selected from random libraries in vitro by using the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technology. Likewise, peptide aptamers are short peptides that can be selected in vitro by using different strategies including phage display, ribosome display, or mRNA display. Aptamers are superior to antibodies with regard to ease of production, high stability, small size, and low cost. Therefore, aptamers find broad use in different biotechnological and therapeutic applications. Among them, aptamer use in virus detection and antiviral therapy is one of the attractive applications. The present Covid-19 pandemic and life-threatening viral infections reveal the need for rapid therapeutic solutions that can efficiently target viral mechanisms. In this respect, the chapter is mainly focused on aptamers with antiviral activity, as well as the use of aptamers in viral detection platforms. First, we summarize aptamer selection technologies that can be performed in vitro. Among them, we briefly explain ribosome display, mRNA display and SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technologies. Then, we review aptamers targeting viral proteins and viral invasion mechanisms. In addition, we give an overview of aptamers developed against viruses. We also discuss the major hurdles in aptamer use, as well as the strategies to improve the drug potential of aptamers. © 2021 Bentham Science Publishers.
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    A review of aptamer-conjugated nanomaterials for analytical sample preparation: Classification according to the utilized nanomaterials
    (Elsevier, 2024) Ugurlu, Ozge; Man, Ezgi; Gok, Oguz; Ulker, Gozde; Soyturk, Hakan; Ozyurt, Canan; Evran, Serap
    Background: Sample extraction before detection is a critical step in analysis. Since targets of interest are often found in complex matrices, the sample can not be directly introduced to the analytical instrument. Nanomaterials with unique physical-chemical properties are excellent supports for use in sorbent-based extraction. However, they lack selectivity and thus need to be functionalized with target-capturing molecules. Antibodies and molecularly imprinted polymers (MIPs) can be used for this purpose, but they have some problems that limit their practical applications. Hence, functionalization of nanomaterials for selectivity remains a problem. Results: Nucleic acid aptamers are affinity reagents that can provide superiority to antibodies since they can be selected in vitro and at a lower cost. Moreover, aptamers can be chemically synthesized and easily modified with different functional groups. Hence, aptamers are good candidates to impart selectivity to the nanomaterials. Recent studies focus on the integration of aptamers with magnetic nanoparticles, carbon-based nanomaterials, metal-organic frameworks, gold nanoparticles, gold nanorods, silica nanomaterials, and nanofibers. The unique properties of nanomaterials and aptamers make the aptamer-conjugated nanomaterials attractive for use in sample preparation. Aptamer-functionalized nanomaterials have been successfully used for selective extraction of proteins, small molecules, and cells from different types of complex samples such as serum, urine, and milk. In particular, magnetic nanoparticles have a wider use due to the rapid extraction of the sample under magnetic field. Significance: In this review, we aim to emphasize how beneficial features of nanomaterials and aptamers could be combined for extraction or enrichment of the analytes from complex samples. We aim to highlight that the benefits are twofold in terms of selectivity and efficiency when employing nanomaterials and aptamers together as a single platform.