RESUMEN
In the fight against cancer, researchers have turned their attention to the eukaryotic initiation factor eIF4E, a protein whose increased level is strongly correlated with the development and progression of various types of cancer. Among the numerous strategies devised to tackle eIF4E overexpression, the use of 5' end mRNA cap analogues has emerged as a promising approach. Here, we present new candidates as potent m7GMP analogues for inhibiting translation and interfacing with eIF4E. By employing an appropriate strategy, we synthesized doubly modified mono- and dinucleotide cap analogues, introducing simultaneous substituents at both the N7 and N2 positions of the guanine ring. This approach was identified as an effective and promising combination. Our findings reveal that these dual modifications increase the potency of the dinucleotide analogue, marking a significant advancement in the development of cancer therapeutics targeting the eIF4E pathway.
RESUMEN
BACKGROUND: A urachal cyst has a rare incidence that has been reported as 1/5,000 live birth. CASE PRESENTATION: We report two patients with a complicated urachal cyst, a 5-year-old female who presented to the emergency department with severe abdominal pain and a 3-year-old female presenting with abdominal pain and constipation. Upon laparoscopic exploration both patients had complicated urachal cysts which were adherent to the urinary bladder. CONCLUSION: Complicated urachal cysts can present with acute abdominal pain.
Asunto(s)
Abdomen Agudo , Laparoscopía , Quiste del Uraco , Femenino , Humanos , Niño , Preescolar , Quiste del Uraco/complicaciones , Quiste del Uraco/diagnóstico , Quiste del Uraco/cirugía , Dolor Abdominal/complicaciones , Abdomen Agudo/etiología , Laparoscopía/efectos adversos , Servicio de Urgencia en HospitalRESUMEN
The ribosome is subjected to post-translational modifications, including phosphorylation, that affect its biological activity. Among ribosomal elements, the P-proteins undergo phosphorylation within the C terminus, the element which interacts with trGTPases or ribosome-inactivating proteins (RIPs); however, the role of phosphorylation has never been elucidated. Here, we probed the function of phosphorylation on the interaction of P-proteins with RIPs using the ribosomal P1-P2 dimer. We determined the kinetic parameters of the interaction with the toxins using biolayer interferometry and microscale thermophoresis. The results present the first mechanistic insight into the function of P-protein phosphorylation, showing that introduction of a negative charge into the C terminus of P1-P2 proteins promotes α-helix formation and decreases the affinity of the P-proteins for the RIPs.
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Fosfoproteínas/química , Fosfoproteínas/metabolismo , Proteínas Ribosómicas/química , Proteínas Ribosómicas/metabolismo , Toxinas Biológicas/metabolismo , Ácido Glutámico/metabolismo , Humanos , Simulación del Acoplamiento Molecular , Fosfoproteínas/genética , Fosforilación , Dominios Proteicos , Proteínas Ribosómicas/genética , Ricina/química , Ricina/metabolismo , Serina/metabolismo , Toxinas Biológicas/química , Tricosantina/química , Tricosantina/metabolismoRESUMEN
Virus-like particles (VLPs), due to their nanoscale dimensions, presence of interior cavities, self-organization abilities and responsiveness to environmental changes, are of interest in the field of nanotechnology. Nevertheless, comprehensive knowledge of VLP self-assembly principles is incomplete. VLP formation is governed by two types of interactions: protein-cargo and protein-protein. These interactions can be modulated by the physicochemical properties of the surroundings. Here, we used brome mosaic virus (BMV) capsid protein produced in an E. coli expression system to study the impact of ionic strength, pH and encapsulated cargo on the assembly of VLPs and their features. We showed that empty VLP assembly strongly depends on pH whereas ionic strength of the buffer plays secondary but significant role. Comparison of VLPs containing tRNA and polystyrene sulfonic acid (PSS) revealed that the structured tRNA profoundly increases VLPs stability. We also designed and produced mutated BMV capsid proteins that formed VLPs showing altered diameters and stability compared to VLPs composed of unmodified proteins. We also observed that VLPs containing unstructured polyelectrolyte (PSS) adopt compact but not necessarily more stable structures. Thus, our methodology of VLP production allows for obtaining different VLP variants and their adjustment to the incorporated cargo.
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Bromovirus/metabolismo , Proteínas de la Cápside/metabolismo , Escherichia coli/metabolismo , Proteínas Recombinantes/metabolismo , Virión/metabolismo , Bromovirus/ultraestructura , Modelos Moleculares , Tamaño de la Partícula , ARN de Transferencia/metabolismo , Temperatura , Virión/ultraestructuraRESUMEN
Protein kinases control diversity of biochemical processes in human organism. Checkpoint 1 kinase (Chk1) is an important element of the checkpoint signalling pathways and is responsible for DNA damage repair. Hence, this kinase plays an essential role in cancer cells survival and has become an important target for anticancer agents. Our previous investigations showed that some arylsulphonyl indazole derivatives displayed anticancer effect in vitro. In the present study, in order to verify possibility of interactions of pyrazole and indazole derivatives with Chk1, we focused on the docking of selected tosyl derivatives of indazole and condensed pyrazole 1-7 to the Chk1 pocket, analysis of interactions involving optimized ligand-protein system using DFT formalism, and estimation of the interaction enthalpy of the ligand-protein complex by applying the PM7 method. The estimation of binding affinity seems to indicate that the indazole 5-substituted with 3,5-dimethylpyrazole 4 and condensed pyrazoloquinoline derivative 7 fit the best to the Chk1-binding pocket. The values of the energy of interaction, i.e. the enthalpy change (ΔHint), were between - 85.06 and - 124.04 kcal mol-1 for the optimized ligand-Chk1 complexes. The relaxation of the ligands within the complexes azole-protein as well as the distribution of hydrogen contacts between the ligands and kinase pocket amino acids was also analysed using molecular dynamics as a supporting method. Graphical Abstract Presentation of methods used to describe the interactions between arylsulphonyl pyrazole derivatives and Chk1 kinase.
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Quinasa 1 Reguladora del Ciclo Celular (Checkpoint 1)/antagonistas & inhibidores , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Pirazoles/farmacología , Humanos , Ligandos , Inhibidores de Proteínas Quinasas/farmacologíaRESUMEN
Herein we describe a synthesis of new isoxazole-containing 5' mRNA cap analogues via a cycloaddition reaction. The obtained analogues show a capability to inhibit cap-dependent translation in vitro and are characterized by a new binding mode in which an isoxazolic ring, instead of guanine, is involved in the stacking effect. Our study provides valuable information toward designing new compounds that can be potentially used as anticancer therapeutics.
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Isoxazoles/química , Isoxazoles/farmacología , Iniciación de la Cadena Peptídica Traduccional/efectos de los fármacos , Análogos de Caperuza de ARN/química , Análogos de Caperuza de ARN/farmacología , Animales , Diseño de Fármacos , Factor 4E Eucariótico de Iniciación/metabolismo , Isoxazoles/síntesis química , Ratones , Simulación del Acoplamiento Molecular , Análogos de Caperuza de ARN/síntesis química , ConejosRESUMEN
Peptides have recently attracted much attention as promising drug candidates. Rational design of peptide-derived therapeutics usually requires structural characterization of the underlying protein-peptide interaction. Given that experimental characterization can be difficult, reliable computational tools are needed. In recent years, a variety of approaches have been developed for 'protein-peptide docking', that is, predicting the structure of the protein-peptide complex, starting from the protein structure and the peptide sequence, including variable degrees of information about the peptide binding site and/or conformation. In this review, we provide an overview of protein-peptide docking methods and outline their capabilities, limitations, and applications in structure-based drug design. Key challenges are also briefly discussed, such as modeling of large-scale conformational changes upon binding, scoring of predicted models, and optimal inclusion of varied types of experimental data and theoretical predictions into an integrative modeling process.
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Diseño de Fármacos , Simulación del Acoplamiento Molecular , Péptidos/farmacología , Animales , Sitios de Unión , Humanos , Péptidos/síntesis química , Unión Proteica , Conformación Proteica , Relación Estructura-ActividadRESUMEN
Activation-induced cytidine deaminase (AID) is known for its established role in antibody production. AID induces the diversification of antibodies by deaminating deoxycytidine (C) within immunoglobulin genes. The capacity of AID to deaminate 5-methyldeoxycytidine (5 mC) and/or 5-hydroxymethyldeoxycytidine (5 hmC), and consequently AID involvement in active DNA demethylation, is not fully resolved. For instance, structural determinants of AID activity on different substrates remain to be identified. To better understand the latter issue, we tested how mutations in human AID (hAID) influence its ability to deaminate C, 5 mC, and 5 hmC in vitro. We showed that each of the selected mutations differentially affects hAID's ability to deaminate C and 5 mC. At the same time, we did not observe hAID activity on 5 hmC. Surprisingly, we found that the N51A hAID mutant, with no detectable activity on C, efficiently deaminated 5 mC, which may suggest different requirements for C and 5 mC deamination. Homology modeling and molecular dynamics simulations revealed that the pattern of enzyme-substrate recognition is one of the important factors determining enzyme activity on C and 5 mC. Consequently, we have proposed mechanisms that explain why wild type hAID more efficiently deaminates C than 5 mC in vitro and why 5 hmC is not deaminated.
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Citidina Desaminasa/metabolismo , Citidina/análogos & derivados , ADN de Cadena Simple , Mutación , Secuencia de Bases , Citidina/metabolismo , Citidina Desaminasa/química , Citidina Desaminasa/genética , Humanos , Metilación , Modelos Moleculares , Conformación ProteicaRESUMEN
Experimental methods based on DNA and RNA hybridization, such as multiplex polymerase chain reaction, multiplex ligation-dependent probe amplification, or microarray analysis, require the use of mixtures of multiple oligonucleotides (primers or probes) in a single test tube. To provide an optimal reaction environment, minimal self- and cross-hybridization must be achieved among these oligonucleotides. To address this problem, we developed EvOligo, which is a software package that provides the means to design and group DNA and RNA molecules with defined lengths. EvOligo combines two modules. The first module performs oligonucleotide design, and the second module performs oligonucleotide grouping. The software applies a nearest-neighbor model of nucleic acid interactions coupled with a parallel evolutionary algorithm to construct individual oligonucleotides, and to group the molecules that are characterized by the weakest possible cross-interactions. To provide optimal solutions, the evolutionary algorithm sorts oligonucleotides into sets, preserves preselected parts of the oligonucleotides, and shapes their remaining parts. In addition, the oligonucleotide sets can be designed and grouped based on their melting temperatures. For the user's convenience, EvOligo is provided with a user-friendly graphical interface. EvOligo was used to design individual oligonucleotides, oligonucleotide pairs, and groups of oligonucleotide pairs that are characterized by the following parameters: (1) weaker cross-interactions between the non-complementary oligonucleotides and (2) more uniform ranges of the oligonucleotide pair melting temperatures than other available software products. In addition, in contrast to other grouping algorithms, EvOligo offers time-efficient sorting of paired and unpaired oligonucleotides based on various parameters defined by the user.
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Sondas de ADN/química , ADN/química , Hibridación de Ácido Nucleico , Oligonucleótidos/química , ARN/química , Programas Informáticos , Algoritmos , HumanosRESUMEN
Mitsunobu reaction of partially acylated uridine proceeds with high regioselectivity for intramolecular SN2 anhydro linkage closuring. Under the reaction conditions, an isomeric mixture of diacyl uridine derivatives with either free 2'- or 3'-hydroxyl group was transformed into a single cyclonucleosidic product, 2,2'-anhydro-3',5'-di-O-acyluridine. This paper presents a possible mechanism of the reactions, the explanation of observed phenomenon based on semiempirical and density functional theory (DFT) calculations and possible utility of this synthetic pathway.
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Uridina/análogos & derivados , Uridina/síntesis química , Acilación , Carbonatos/síntesis química , Conformación Molecular , TermodinámicaRESUMEN
1,25-dihydroxyvitamin D(3) has quite significant anticancer properties, but its strong calcemic effect in principle excludes it as a potential anticancer drug. Currently, a lot of effort is being devoted to develop potent anticancer analogs of 1,25-dihydroxyvitamin D(3) that would not induce hypercalcemia during therapy. In this work, the free binding energy of the VDR receptor with 1,25-dihydroxyvitamin D(3) and its three potent analogs (EB 1089, KH 1060 and RO 25-9022) is calculated and compared with each other. With this approach, we could estimate the relative binding affinity of the most potent analog, RO 25-9022, and also revealed a quite distinct mechanism of its interaction with VDR.
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Antineoplásicos , Calcitriol , Conformación Proteica , Receptores de Calcitriol/química , Antineoplásicos/química , Antineoplásicos/farmacología , Sitios de Unión , Calcitriol/análogos & derivados , Calcitriol/síntesis química , Calcitriol/química , Línea Celular Tumoral , Cristalografía por Rayos X , Metabolismo Energético , Humanos , Hipercalcemia/inducido químicamente , Hipercalcemia/complicaciones , Ligandos , Simulación del Acoplamiento Molecular , Neoplasias/tratamiento farmacológico , Unión Proteica , Receptores de Calcitriol/metabolismoRESUMEN
Understanding the interactions of epothilones with ß-tubulin is crucial for computer aided rational design of macrocyclic drugs based on epothilones and epothilone derivatives. Despite numerous structure-activity relationship investigations we still lack substantial knowledge about the binding mode of epothilones and their derivatives to ß-tubulin. In this work, we reevaluated the electron crystallography structure of epothilone A/ß-tubulin complex (PDB entry 1TVK) and proposed an alternative binding mode of epothilone A to ß-tubulin that explains more experimental facts.