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Order Tymovirales
Published in Paul Pumpens, Peter Pushko, Philippe Le Mercier, Virus-Like Particles, 2022
Paul Pumpens, Peter Pushko, Philippe Le Mercier
To develop this approach further, Dickmeis et al. (2015) created a novel production system for the mosaic PVX particles, after the previously described 2A-based approach. The different polypeptides were displayed as the coat fusions when combinations of PVX and tobacco mosaic (TMV) expression vectors were used, each expressing different PVX coat fusions. To prove the principle of the assembly of mosaic chimeric PVX virions, the GFP and the red fluorescent protein mCherry were chosen, as well as a bimolecular fluorescence complementation (BiFC) system with split-mCherry as coat fusion proteins. The presence of assembled split-mCherry on the surface confirmed the mosaic character of the chimeric particles (Dickmeis et al. 2015).
Non-VLPs
Published in Paul Pumpens, Single-Stranded RNA Phages, 2020
The MS2-based platform contributed to the development of novel imaging tools, such as far-red mNeptune-based bimolecular fluorescence complementation (BiFC), and TriFC systems with excitation and emission above 600 nm in the “tissue optical window” for imaging of protein–protein and RNA-protein interactions in live cells and mice (Han Y et al. 2014).
Targeting protein-protein interactions with low molecular weight and short peptide modulators: insights on disease pathways and starting points for drug discovery
Published in Expert Opinion on Drug Discovery, 2023
Daniela Trisciuzzi, Bruno O. Villoutreix, Lydia Siragusa, Massimo Baroni, Gabriele Cruciani, Orazio Nicolotti
Some experimental/computational technologies are used to quantify the detection of transient PPIs [111]. For instance, yeast two-hybrid (Y2H) is a well-known method for large-scale detection of PPIs although in the case of weak transient interactions (Kd ~ 10−6M), a high number of false-positive cases have been observed. The use of chemical crosslinking technologies can provide additional information by freezing transiently formed complexes through covalent-bond formation between the interacting partners. In addition, the implementation of crosslink during the washing phase of TAP-MS enhances the ability of this method to detect transient interactions. The bimolecular fluorescence complementation (BiFC) assays are able to detect transient (as well as permanent) PPIs in intact cells bypassing the purification phases. Computational approaches are also widely useful to investigate the affinity of PPIs as less costly and labor-intensive than experimental methods [109]. For instance, bioinformatics tools able to identify co-conserved and co-expressed genes with the use of statistical framework seem well suited to predict strongly correlated stable complexes and transient interactions.
Targeting the intrinsically disordered architectural High Mobility Group A (HMGA) oncoproteins in breast cancer: learning from the past to design future strategies
Published in Expert Opinion on Therapeutic Targets, 2020
Silvia Pegoraro, Gloria Ros, Michela Sgubin, Sara Petrosino, Alberto Zambelli, Riccardo Sgarra, Guidalberto Manfioletti
b) New directions to interfere with HMGA activities: The protein/protein interaction aspect of HMGA proteins has not yet been deeply addressed. HMGA proteins have been demonstrated to be hubs in the chromatin network and to be involved in a large number of protein/protein (p/p) interactions [23,24,188], for most of which a functional outcome has been provided [19]. Why have HMGA p/p interactions not been explored as potential drug targets? In our opinion, the answer relies on the ID status of HMGA proteins. HMGA proteins have a peculiar structural organization and amino acid composition that make them prototypes for the IDP category [22]. Structural-based approaches are currently being used for protein-binding drug discovery, in which the structural features of proteins constitute guides for the selection of potential binders. Clearly, these strategies cannot be applied in the absence of a fixed structure. There is a growing interest in IDPs since it was realized that IDPs are indeed widely involved in many pathological conditions, often with a causal role [189]. Indeed, it would be worth investigating large-scale screening to select for HMGA binders. Even if some effort has been made to pave the way toward rational drug design strategies against IDPs, the main successful approaches have made use of high-throughput molecular screening [190]. The technological platforms to perform this kind of investigation are already available and span from in vivo bimolecular fluorescence complementation (BiFC) assays to the more automated in vitro high-throughput surface plasmon resonance (HT-SPR) screenings [191,192].
Molecular mechanisms governing axonal transport: a C. elegans perspective
Published in Journal of Neurogenetics, 2020
Amruta Vasudevan, Sandhya P. Koushika
In vitro measurements of run lengths of Kinesin-3 motors are much shorter (∼1-10µm) than cargo run lengths observed in vivo (∼15–30 µm), which can be attributed to the presence of factors that function to enhance the motility characteristics of molecular motors in vivo (Beeg et al., 2008; Sood et al., 2018; Verbrugge, van den Wildenberg & Peterman, 2009). Some of these factors, identified from mammalian and C. elegans studies, have been listed in Table 1. Investigations conducted in C. elegans neurons revealed that SYD-2/Liprin-α (Wagner et al., 2009) and LIN-2 (a MAGUK protein containing a CaMKII-homologous domain) function to cluster and increase the processive motility and velocity of UNC-104 (Figure 1(b)) (Wu, Muthaiyan Shanmugam, Bhan, Huang, & Wagner, 2016). CASY-1 (a calsyntenin family protein) was discovered to regulate UNC-104-mediated synaptic vesicle transport specifically in GABAergic motor neurons, likely by enhancing the motility of UNC-104 (Thapliyal et al., 2018), thereby promoting anterograde transport. Bimolecular Fluorescence Complementation (BiFC), a technique that tests for the direct interaction between two proteins by fusing one to the N-terminal fragment and the other to the C-terminal fragment of Venus fluorescent protein and examining reconstituted fluorescence, identified that neuronal adaptors and interactors of UNC-104 regulate its subcellular distribution (Hsu, Moncaleano, & Wagner, 2011). UNC-104 bound to UNC-16/JIP-3 predominantly localizes to the neuronal cell soma, while UNC-104 bound to DNC-1, a component of the dynactin complex, largely localizes to the distal tips of neurons, and UNC-104 bound to SYD-2/Liprin-α distributes along the neuronal process (Hsu et al., 2011). These interactions have been suggested to influence the velocity and persistence times of UNC-104, with implications for axonal transport of synaptic vesicles (Hsu et al., 2011).