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1.
Molecules ; 25(1)2019 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-31877731

RESUMO

There is a continuous search for more reliable and effective alternatives to control phytopathogens through different strategies. In this context, indole-containing phytoalexins are stimuli-induced compounds implicated in plant defense against plant pathogens. However, phytoalexins' efficacy have been limited by fungal detoxifying mechanisms, thus, the research on bioisosteres-based analogs can be a friendly alternative regarding the control of Fusarium phytopathogens, but there are currently few studies on it. Thus, as part of our research on antifungal agents, a set of 21 synthetic indole-containing phytoalexin analogs were evaluated as inhibitors against the phyopathogen Fusarium oxysporum. Results indicated that analogs of the N,N-dialkylthiourea, N,S-dialkyldithiocarbamate and substituted-1,3-thiazolidin-5-one groups exhibited the best docking scores and interaction profiles within the active site of Fusarium spp. enzymes. Vina scores exhibited correlation with experimental mycelial growth inhibition using supervised statistics, and this antifungal dataset correlated with molecular interaction fields after CoMFA. Compound 24 (tert-butyl (((3-oxo-1,3-diphenylpropyl)thio)carbonothioyl)-l-tryptophanate), a very active analog against F. oxysporum, exhibited the best interaction with lanosterol 14α-demethylase according to molecular docking, molecular dynamics and molecular mechanic/poisson-boltzmann surface area (MM/PBSA) binding energy performance. After data analyses, information on mycelial growth inhibitors, structural requirements and putative enzyme targets may be used in further antifungal development based on phytoalexin analogs for controlling phytopathogens.


Assuntos
Antifúngicos/síntese química , Fusarium/crescimento & desenvolvimento , Indóis/síntese química , Sesquiterpenos/química , Antifúngicos/química , Antifúngicos/farmacologia , Domínio Catalítico/efeitos dos fármacos , Simulação por Computador , Enzimas/química , Enzimas/farmacologia , Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Fusarium/efeitos dos fármacos , Fusarium/enzimologia , Indóis/química , Indóis/farmacologia , Viabilidade Microbiana/efeitos dos fármacos , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Estrutura Molecular
2.
PLoS One ; 14(11): e0224132, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31697708

RESUMO

Succinate dehydrogenase (SDH) inhibitors (SDHIs) are used worldwide to limit the proliferation of molds on plants and plant products. However, as SDH, also known as respiratory chain (RC) complex II, is a universal component of mitochondria from living organisms, highly conserved through evolution, the specificity of these inhibitors toward fungi warrants investigation. We first establish that the human, honeybee, earthworm and fungal SDHs are all sensitive to the eight SDHIs tested, albeit with varying IC50 values, generally in the micromolar range. In addition to SDH, we observed that five of the SDHIs, mostly from the latest generation, inhibit the activity of RC complex III. Finally, we show that the provision of glucose ad libitum in the cell culture medium, while simultaneously providing sufficient ATP and reducing power for antioxidant enzymes through glycolysis, allows the growth of RC-deficient cells, fully masking the deleterious effect of SDHIs. As a result, when glutamine is the major carbon source, the presence of SDHIs leads to time-dependent cell death. This process is significantly accelerated in fibroblasts derived from patients with neurological or neurodegenerative diseases due to RC impairment (encephalopathy originating from a partial SDH defect) and/or hypersensitivity to oxidative insults (Friedreich ataxia, familial Alzheimer's disease).


Assuntos
Transporte de Elétrons/efeitos dos fármacos , Praguicidas/farmacologia , Succinato Desidrogenase/antagonistas & inibidores , Animais , Antioxidantes/metabolismo , Abelhas/metabolismo , Células Cultivadas , Farmacorresistência Fúngica/efeitos dos fármacos , Proteínas Fúngicas/farmacologia , Fungos/metabolismo , Humanos , Membranas Mitocondriais/efeitos dos fármacos , Doenças Neurodegenerativas/tratamento farmacológico , Doenças Neurodegenerativas/metabolismo , Oligoquetos/metabolismo , Succinato Desidrogenase/metabolismo
3.
ACS Appl Mater Interfaces ; 11(43): 39436-39448, 2019 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-31580644

RESUMO

Laccases are multi-copper oxidase enzymes having widespread applications in various biotechnological fields. However, low stability of free enzymes restricts their industrial use. Development of effective methods to preserve and even increase the enzymatic activity is critical to maximize their use, though this remains a challenge. In the present study we immobilized Trametes versicolor laccase on pH-responsive (and charge-switchable) Pluronic-stabilized silver nanoparticles (AgNPsTrp). Our results demonstrate that colloidal stabilization of AgNPsTrp with the amphiphilic copolymer Pluronic F127 enhances enzyme activity (AgNPsTrpF1 + Lac6) by changing the active site microenvironment, which is confirmed by circular dichroism (CD) and fluorescence spectroscopy. Detailed kinetic and thermodynamic studies reveal a facile strategy to improve the protein quality by lowering the activation energy and expanding the temperature window for substrate hydrolysis. The immobilized nanocomposite did not show any change in flow behavior which indirectly suggests that the enzyme stability is maintained, and the enzyme did not aggregate or unfold upon immobilization. Finally, assessing the anticancer efficacy of this nanocomposite in breast cancer MCF-7 cells shows the inhibition of cell proliferation through ß-estradiol degradation and cells apoptosis. To understand the molecular mechanism involved in this process, semi qRT-PCR experiments were performed, which indicated significant decrease in the mRNA levels of anti-apoptotic genes, for example, BCL-2 and NF-kß, and increase in the mRNA level of pro-apoptotic genes like p53 in treated cells, compared to control. Overall, this study offers a completely new strategy for tailoring nano-bio-interfaces with improved activity and stability of laccase.


Assuntos
Neoplasias da Mama , Enzimas Imobilizadas , Proteínas Fúngicas , Lacase , Poloxâmero , Polyporaceae/enzimologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Estabilidade Enzimática , Enzimas Imobilizadas/química , Enzimas Imobilizadas/farmacologia , Feminino , Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Lacase/química , Lacase/farmacologia , Células MCF-7 , Proteínas de Neoplasias/biossíntese , Poloxâmero/química , Poloxâmero/farmacologia
4.
Int J Mol Sci ; 20(21)2019 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-31661772

RESUMO

Our previous research has shown that a fungal immunomodulatory protein from Nectria haematococca (FIP-nha) possesses a wide spectrum of anti-tumor activities, and FIP-nha induced A549 apoptosis by negatively regulating the PI3K/Akt signaling pathway based on comparative quantitative proteomics. This study further confirmed that the anti-lung cancer activity of FIP-nha was significantly stronger than that of the reported LZ-8 and FIP-fve. Subsequently, 1H NMR-based metabolomics was applied to comprehensively investigate the underlying mechanism, and a clear separation of FIP-nha-treated and untreated groups was achieved using pattern recognition analysis. Four potential pathways associated with the anti-tumor effect of FIP-nha on A549 cells were identified, and these were mainly involved in glycolysis, taurine and hypotaurine metabolism, fructose and mannose metabolism, and glycerolipid metabolism. Metabolic pathway analysis demonstrated that FIP-nha could induce A549 cell apoptosis partly by regulating the p53 inhibition pathway, which then disrupted the Warburg effect, as well as through other metabolic pathways. Using RT-PCR analysis, FIP-nha-induced apoptosis was confirmed to occur through upregulation of p53 expression. This work highlights the possible use of FIP-nha as a therapeutic adjuvant for lung cancer treatment.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Proteínas Fúngicas/farmacologia , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/metabolismo , Nectria , Proteína Supressora de Tumor p53/metabolismo , Células A549 , Antineoplásicos/uso terapêutico , Vias Biossintéticas , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/uso terapêutico , Células HEK293 , Humanos , Fosfatidilinositol 3-Quinases/metabolismo , Proteômica , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/genética
5.
Proc Natl Acad Sci U S A ; 116(44): 22353-22358, 2019 10 29.
Artigo em Inglês | MEDLINE | ID: mdl-31611414

RESUMO

An Australian estuarine isolate of Penicillium sp. MST-MF667 yielded 3 tetrapeptides named the bilaids with an unusual alternating LDLD chirality. Given their resemblance to known short peptide opioid agonists, we elucidated that they were weak (K i low micromolar) µ-opioid agonists, which led to the design of bilorphin, a potent and selective µ-opioid receptor (MOPr) agonist (K i 1.1 nM). In sharp contrast to all-natural product opioid peptides that efficaciously recruit ß-arrestin, bilorphin is G protein biased, weakly phosphorylating the MOPr and marginally recruiting ß-arrestin, with no receptor internalization. Importantly, bilorphin exhibits a similar G protein bias to oliceridine, a small nonpeptide with improved overdose safety. Molecular dynamics simulations of bilorphin and the strongly arrestin-biased endomorphin-2 with the MOPr indicate distinct receptor interactions and receptor conformations that could underlie their large differences in bias. Whereas bilorphin is systemically inactive, a glycosylated analog, bilactorphin, is orally active with similar in vivo potency to morphine. Bilorphin is both a unique molecular tool that enhances understanding of MOPr biased signaling and a promising lead in the development of next generation analgesics.


Assuntos
Analgésicos Opioides/farmacologia , Proteínas Fúngicas/farmacologia , Oligopeptídeos/farmacologia , Penicillium/química , Receptores Opioides mu/agonistas , Analgésicos Opioides/química , Animais , Sítios de Ligação , Linhagem Celular Tumoral , Proteínas Fúngicas/química , Células HEK293 , Humanos , Camundongos , Simulação de Acoplamento Molecular , Oligopeptídeos/química , Ligação Proteica , Receptores Opioides mu/química , Receptores Opioides mu/metabolismo
6.
Chem Pharm Bull (Tokyo) ; 67(10): 1023-1029, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31582623

RESUMO

The combinatorial synthesis and biological evaluation of destruxins are described herein. First, the total synthesis of destruxin E was achieved, and its absolute configuration was successfully determined to be (S). In addition, the preparation of a combinatorial library based on the structure of destruxins was carried out by the split-and-pool method. Biological evaluation of the resulting analogs against osteoclast-like multinuclear cells (OCLs) revealed that the N-methyl-alanine residue was crucial to inducing morphological changes in OCLs. In particular, functionalization at the ß-position of the proline (Pro) residue was found to be tolerant of the desired biological activity of destruxin E, suggesting that the ß-position of the Pro residue should be a promising site for the introduction of a chemical tag toward the preparation of a molecular probe.


Assuntos
Depsipeptídeos/farmacologia , Proteínas Fúngicas/farmacologia , Osteoclastos/efeitos dos fármacos , Técnicas de Química Combinatória , Depsipeptídeos/síntese química , Depsipeptídeos/química , Proteínas Fúngicas/síntese química , Proteínas Fúngicas/química , Humanos , Estrutura Molecular , Relação Estrutura-Atividade
7.
Int J Biol Macromol ; 141: 517-528, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31494159

RESUMO

Chitin, a crucial structural and functional component of insects and fungi, serves as a target for pest management by utilizing novel chitinases. Here, we report the biocontrol potential of recombinant Myrothecium verrucaria endochitinase (rMvEChi) against insect pest and fungal pathogens. A complete ORF of MvEChi (1185 bp) was cloned and heterologously expressed in Escherichia coli. Structure based sequence alignment of MvEChi revealed the presence of conserved domains SXGG and DXXDXDXE specific for GH-18 family, involved in substrate binding and catalysis, respectively. rMvEChi (46.6 kDa) showed optimum pH and temperature as 7.0 and 30 °C, respectively. Furthermore, rMvEChi remained stable within the pH range of 6.0 to 8.0 and up to 40 °C. rMvEChi exhibited kcat/Km values of 129.83 × 103 [(g/L)-1 s-1] towards 4MU chitotrioside. Hydrolysis of chitooligosaccharides with various degrees of polymerization (DP) using rMvEChi indicated the release of DP2 as main end product with order of reaction as DP6 > DP5 > DP4 > DP3. Bioassay of rMvEChi against Helicoverpa armigera displayed potent anti-feedant activity and induced mortality. In vitro antifungal activity against plant pathogenic fungi (Ustilago maydis and Bipolaris sorokiniana) exhibited significant inhibition of mycelium growth. These results suggest that MvEChi has significant potential in enzyme-based pest and pathogen management.


Assuntos
Ascomicetos/enzimologia , Quitinases , Proteínas Fúngicas , Lepidópteros/crescimento & desenvolvimento , Doenças das Plantas/microbiologia , Ustilago/crescimento & desenvolvimento , Animais , Quitinases/química , Quitinases/farmacologia , Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Proteínas Recombinantes/química , Proteínas Recombinantes/farmacologia
8.
Food Microbiol ; 84: 103266, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421779

RESUMO

Postharvest fungal diseases are among the main causes of fresh fruit losses. Chemical control is against claims for "natural" or "chemical-free" products. Biocontrol agents, such as antifungal proteins or their producing moulds, may serve to combat unwanted pathogens. Since the effectiveness of these bioprotective agents depends on the food substrate, their effect must be tested on fruits. The objective of this work was to study the effect of the antifungal protein PgAFP and its producer, Penicillium chrysogenum, against Penicillium expansum and Penicillium digitatum growth on apple and oranges respectively, and the PgAFP effect on eleven P. expansum, Penicillium italicum, and P. digitatum strains in vitro, and on patulin production on apple substrate. The sensitivity upon PgAFP was P. digitatum > P. expansum > P. italicum. In oranges, broadly, no inhibitory effect was obtained. PgAFP and P. chrysogenum did not inhibit the P. expansum CMP-1 growth on Golden Delicious apples, however, a successful effect was achieved on Royal Gala apples. On apple substrate, patulin production by P. expansum CMP-1 rose in parallel to PgAFP concentrations, linked with high reactive oxygen species levels. PgAFP cannot be proposed as a bioprotective agent on apple. However, P. chrysogenum is a promising agent to be used on Royal Gala apples.


Assuntos
Antifúngicos/farmacologia , Citrus/microbiologia , Proteínas Fúngicas/farmacologia , Malus/microbiologia , Penicillium chrysogenum/química , Penicillium/efeitos dos fármacos , Microbiologia de Alimentos , Proteínas Fúngicas/química , Patulina/biossíntese
9.
BMC Res Notes ; 12(1): 471, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31370903

RESUMO

OBJECTIVES: Tan spot is a yield-reducing disease that affects wheat and is caused by the fungus Pyrenophora tritici-repentis (Ptr). Eight races of Ptr have been identified based upon production of the effectors Ptr ToxA, Ptr ToxB, and Ptr ToxC. Wheat cultivars have also been characterized by their resistance and susceptibility to races of Ptr and sensitivity to the effectors. The objective of this research was to assess differences in gene expression between Ptr resistant and susceptible wheat cultivars when either inoculated with Ptr race 2 spores or directly infiltrated with Ptr ToxA. DATA DESCRIPTION: A greenhouse experiment was used to assess wheat-Ptr interaction. Wheat seedlings were grown for two weeks prior to the experiment under greenhouse conditions. Four treatments were used: (1) spray-inoculation with a suspension of Ptr spores (3000 spores/mL) (2) spray inoculation with water as a control (3) needleless syringe injection with Ptr ToxA, and (4) needleless syringe injection with water as a control. Plants were transferred to a humidity chamber and leaf sample were taken at 0, 8, and 16 h. After RNA extraction and sequencing, 48 RNA datasets are reported. This data will be useful in understanding how resistant wheat responds to Ptr compared to susceptible wheat.


Assuntos
Doenças das Plantas/genética , Folhas de Planta/genética , RNA de Plantas/genética , Saccharomycetales/patogenicidade , Transcriptoma , Triticum/genética , Resistência à Doença/genética , Suscetibilidade a Doenças , Proteínas Fúngicas/farmacologia , Interações Hospedeiro-Patógeno/genética , Micotoxinas/farmacologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/imunologia , Folhas de Planta/microbiologia , Saccharomycetales/fisiologia , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/imunologia , Plântula/microbiologia , Esporos Fúngicos/patogenicidade , Esporos Fúngicos/fisiologia , Triticum/efeitos dos fármacos , Triticum/imunologia , Triticum/microbiologia
10.
Int J Biol Macromol ; 134: 1097-1107, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31103591

RESUMO

Lectins are unique biorecognition molecules that identify cell surface carbohydrates/glycoproteins and play significant role in various interactions. They are ubiquitous glycan-specific proteins/glycoproteins of non-immune origin. Amongst microfungi, lectins have been widely reported from aspergilli, penicilli, Fusarium sp., etc., however a plethora of genera still remains unexplored. Microfungal lectins have wide diversity in their haemagglutination and carbohydrate specificity. They also exhibit great variations in their structural organization which influences lectin-glycan interactions. The present review summarizes the sources, characteristics and structural diversity of microfungal lectins. Prospective biomedical applications of microfungal lectins as anticancer, mitogenic, immunomodulatory, antioxidant and therapeutic agents have been discussed extensively.


Assuntos
Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Lectinas/química , Lectinas/farmacologia , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Humanos , Modelos Moleculares , Conformação Proteica , Relação Estrutura-Atividade
11.
Nutrients ; 11(4)2019 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-30934760

RESUMO

Frailty is a geriatric syndrome associated with both locomotor and cognitive decline, implicated in both poor quality of life and negative health outcomes. One central question surrounding frailty is whether phenotypic frailty is associated with the cognitive impairment during aging. Using spontaneous behavioral tests and by studying the dynamic change during aging, we demonstrated that the two form of vulnerability, locomotor and recognition memory decline, develop in parallel and therefore, integration of the motoric and cognitive evaluations are imperative. We developed an integrated frailty index based on both phenotypic and recognition memory performances. Hericium erinaceus (H. erinaceus) is a medicinal mushroom that improves recognition memory in mice. By using HPLC-UV-ESI/MS analyses we obtained standardized amounts of erinacine A and hericenones C and D in H. erinaceus extracts, that were tested in our animal model of physiological aging. Two-month oral supplementation with H. erinaceus reversed the age-decline of recognition memory. Proliferating cell nuclear antigen (PCNA) and doublecortin (DCX) immunohistochemistry in the hippocampus and cerebellum in treated mice supported a positive effect of an H. erinaceus on neurogenesis in frail mice.


Assuntos
Agaricales , Fragilidade/tratamento farmacológico , Proteínas Fúngicas/farmacologia , Neurogênese/efeitos dos fármacos , Reconhecimento Psicológico/efeitos dos fármacos , Envelhecimento/efeitos dos fármacos , Animais , Cerebelo/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Camundongos
12.
Braz J Microbiol ; 50(3): 603-612, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30982213

RESUMO

Herein, the class II hydrophobin gene HFBII-4 was cloned from the biocontrol agent Trichoderma asperellum ACCC30536 and recombinant rHFBII-4 was expressed in Pichia pastoris GS115. Treatment of Populus davidiana × P. alba var. pyramidalis (PdPap poplar) with rHFBII-4 altered the expression levels of genes in the auxin, salicylic acid (SA), and jasmonic acid (JA) signal transduction pathways. Polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) enzyme activities were induced with rHFBII-4. Evans Blue and nitro blue tetrazolium (NBT) staining indicated that cell membrane permeability and reactive oxygen species were lower in the leaves of plants treated with rHFBII-4. The chlorophyll content was higher than that of control at 2-5 days after treatment. Furthermore, poplar seedlings were inoculated with Alternaria alternata, disease symptoms were observed. The diseased area was smaller in leaves induced with rHFBII-4 compared with control. In summary, rHFBII-4 enhances resistance to A. alternata.


Assuntos
Proteínas Fúngicas/farmacologia , Doenças das Plantas/microbiologia , Populus/efeitos dos fármacos , Populus/imunologia , Trichoderma/metabolismo , Alternaria/fisiologia , Ciclopentanos/imunologia , Resistência à Doença , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Oxilipinas/imunologia , Doenças das Plantas/imunologia , Reguladores de Crescimento de Planta/imunologia , Folhas de Planta/imunologia , Folhas de Planta/microbiologia , Populus/microbiologia , Trichoderma/química , Trichoderma/genética
13.
BMC Genomics ; 20(1): 243, 2019 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-30909884

RESUMO

BACKGROUND: Fungi are an attractive source of nutrients for predators. As part of their defense, some fungi are able to induce the production of anti-predator protein toxins in response to predation. A previous study on the interaction of the model mushroom Coprinopsis cinerea by the fungivorous nematode Aphelenchus avenae on agar plates has shown that the this fungal defense response is most pronounced in the part of the mycelium that is in direct contact with the nematode. Hence, we hypothesized that, for a comprehensive characterization of this defense response, an experimental setup that maximizes the zone of direct interaction between the fungal mycelium and the nematode, was needed. RESULTS: In this study, we conducted a transcriptome analysis of C. cinerea vegetative mycelium upon challenge with A. avenae using a tailor-made microfluidic device. The device was designed such that the interaction between the fungus and the nematode was confined to a specific area and that the mycelium could be retrieved from this area for analysis. We took samples from the confrontation area after different time periods and extracted and sequenced the poly(A)+ RNA thereof. The identification of 1229 differentially expressed genes (DEGs) shows that this setup profoundly improved sensitivity over co-cultivation on agar plates where only 37 DEGs had been identified. The product of one of the most highly upregulated genes shows structural homology to bacterial pore-forming toxins, and revealed strong toxicity to various bacterivorous nematodes. In addition, bacteria associated with the fungivorous nematode A. avenae were profiled with 16S rRNA deep sequencing. Similar to the bacterivorous and plant-feeding nematodes, Proteobacteria and Bacteroidetes were the most dominant phyla in A. avenae. CONCLUSIONS: The use of a novel experimental setup for the investigation of the defense response of a fungal mycelium to predation by fungivorous nematodes resulted in the identification of a comprehensive set of DEGs and the discovery of a novel type of fungal defense protein against nematodes. The bacteria found to be associated with the fungivorous nematode are a possible explanation for the induction of some antibacterial defense proteins upon nematode challenge.


Assuntos
Agaricales/imunologia , Proteínas Fúngicas/genética , Técnicas Analíticas Microfluídicas/métodos , Nematoides/patogenicidade , Análise de Sequência de RNA/métodos , Agaricales/genética , Animais , Bacteroides/genética , Bacteroides/isolamento & purificação , Proteínas Fúngicas/farmacologia , Perfilação da Expressão Gênica/métodos , Regulação Fúngica da Expressão Gênica , Nematoides/efeitos dos fármacos , Nematoides/microbiologia , Filogenia , Proteobactérias/genética , Proteobactérias/isolamento & purificação , RNA Ribossômico 16S/genética
14.
Nanoscale ; 11(7): 3248-3260, 2019 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-30706922

RESUMO

The development of drug delivery and imaging tools is a major challenge in human health, in particular in cancer pathologies. This work describes the optimization of a protein nanocontainer, belonging to the lectin protein family, for its use in epithelial cancer diagnosis and treatment. Indeed, it specifically targets a glycosidic marker, the T antigen, which is known to be characteristic of epithelial cancers. Its quaternary structure reveals a large hydrated inner cavity able to transport small therapeutic molecules. Optimization of the nanocontainer by site directed mutagenesis allowed controlling loading and release of confined drugs. Doxorubicin confinement was followed, both theoretically and experimentally, and provided a proof of concept for the use of this nanocontainer as a vectorization system. In OVCAR-3 cells, a human ovarian adenocarcinoma cell line that expresses the T antigen, the drug was observed to be delivered inside late endosomes/lysosomes. These results show that this new type of vectorization and imaging device opens new exciting perspectives in nano-theranostic approaches.


Assuntos
Adenocarcinoma/tratamento farmacológico , Antígenos Virais de Tumores/metabolismo , Doxorrubicina , Sistemas de Liberação de Medicamentos , Proteínas Fúngicas , Lectinas , Nanoestruturas , Neoplasias Ovarianas/tratamento farmacológico , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Basidiomycota/genética , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Feminino , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/farmacologia , Humanos , Lectinas/química , Lectinas/genética , Lectinas/farmacologia , Nanoestruturas/química , Nanoestruturas/uso terapêutico , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia
15.
Peptides ; 113: 52-65, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30738838

RESUMO

Lipopeptide antibiotics have linear or cyclic structures with one or more hydrocarbon tails linked to the N-terminus of a short oligopeptide that may be chemically modified and/or contain unusual amino acid residues in their structures. They possess huge potential as pharmaceutical drugs and biocontrol agents, and ˜30 representative genera of fungi are known to produce them. Some chemically synthesised derivatives have already been developed into commercial products or subjected to clinical trials, including cilofungin, caspofungin, micafungin, anidulafungin, rezafungin, emodepside, fusafungine and destruxins. This review summarizes 200 fungi-derived compounds reported since 2000, including 95 cyclic depsipeptides, 67 peptaibiotics (including 35 peptaibols, eight lipoaminopeptides, and five lipopeptaibols), and 38 non-depsipeptide and non-peptaibiotic lipopeptides. Their sources, structural sequences, antibiotic activities (e.g. antibacterial, antifungal, antiviral, antimycobacterial, antimycoplasmal, antimalarial, antileishmanial, insecticidal, antitrypanosomal and nematicidal), structure-activity relationships, mechanisms of action, and specific relevance are discussed. These compounds have attracted considerable interest within the pharmaceutical and agrochemical industries.


Assuntos
Anti-Infecciosos/farmacologia , Lipopeptídeos/farmacologia , Anidulafungina/farmacologia , Caspofungina/farmacologia , Depsipeptídeos/farmacologia , Equinocandinas/farmacologia , Proteínas Fúngicas/farmacologia , Fungos/química , Micafungina/farmacologia , Relação Estrutura-Atividade
16.
Mol Nutr Food Res ; 63(7): e1801231, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30667154

RESUMO

SCOPE: The prevalence of obesity and related disorders has vastly increased throughout the world and prevention of such circumstances thus represents a major challenge. Here, it has been shown that one protein-bound ß-glucan (PBG) from the edible mushroom Coriolus versicolor can be a potent anti-obesity component. METHODS AND RESULTS: PBG can reduce obesity and metabolic inflammation in mice fed with a high-fat diet (HFD). Gut microbiota analysis reveals that PBG markedly increases the abundance of Akkermansia muciniphila, although it does not rescue HFD-induced change in the Firmicutes to Bacteroidetes ratio. It appears that PBG alters host physiology and creates an intestinal microenvironment favorable for A. muciniphila colonization. Fecal transplants from PBG-treated animals in part reduce obesity in recipient HFD-fed mice. Further, PBG is shown to upregulate expression of a set of genes related to host metabolism in microbiota-depleted mice. CONCLUSION: The data highlight that PBG may exert its anti-obesity effects through a mirobiota-dependent (richness of specific microbiota) and -independent (modulation of host metabolism) manner. The fact that C. versicolor PBGs are approved oral immune boosters in cancers and chronic hepatitis with well-established safety profiles may accelerate PBG as a novel use for obesity treatment.


Assuntos
Agaricales/química , Fármacos Antiobesidade/farmacologia , Obesidade/prevenção & controle , beta-Glucanas/química , beta-Glucanas/farmacologia , Animais , Fármacos Antiobesidade/química , Citocinas/sangue , Dieta Hiperlipídica/efeitos adversos , Avaliação Pré-Clínica de Medicamentos/métodos , Transplante de Microbiota Fecal , Feminino , Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Camundongos Endogâmicos C57BL , Obesidade/etiologia , Obesidade/terapia , Verrucomicrobia/efeitos dos fármacos
17.
Structure ; 27(3): 449-463.e7, 2019 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-30595457

RESUMO

Hsp104 is an AAA+ protein disaggregase with powerful amyloid-remodeling activity. All nonmetazoan eukaryotes express Hsp104 while eubacteria express an Hsp104 ortholog, ClpB. However, most studies have focused on Hsp104 from Saccharomyces cerevisiae and ClpB orthologs from two eubacterial species. Thus, the natural spectrum of Hsp104/ClpB molecular architectures and protein-remodeling activities remains largely unexplored. Here, we report two structures of Hsp104 from the thermophilic fungus Calcarisporiella thermophila (CtHsp104), a 2.70Å crystal structure and 4.0Å cryo-electron microscopy structure. Both structures reveal left-handed, helical assemblies with all domains clearly resolved. We thus provide the highest resolution and most complete view of Hsp104 hexamers to date. We also establish that CtHsp104 antagonizes several toxic protein-misfolding events in vivo where S. cerevisiae Hsp104 is ineffective, including rescue of TDP-43, polyglutamine, and α-synuclein toxicity. We suggest that natural Hsp104 variation is an invaluable, untapped resource for illuminating therapeutic disaggregases for fatal neurodegenerative diseases.


Assuntos
Adenosina Trifosfatases/química , Adenosina Trifosfatases/farmacologia , Mucorales/enzimologia , Microscopia Crioeletrônica , Cristalografia por Raios X , Proteínas de Ligação a DNA/antagonistas & inibidores , Proteínas Fúngicas/química , Proteínas Fúngicas/farmacologia , Humanos , Modelos Moleculares , Peptídeos/antagonistas & inibidores , Conformação Proteica em alfa-Hélice , Deficiências na Proteostase/prevenção & controle , alfa-Sinucleína/antagonistas & inibidores
18.
World J Microbiol Biotechnol ; 35(2): 27, 2019 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-30680515

RESUMO

In this study, the cDNA of immunomodulatory protein from Poria cocos (PCP) was amplified by reverse transcription polymerase chain reaction and used to transform P. Pastoris cells, resulting in rPCP expression as a secreted protein to a concentration of ~ 38 mg/L following methanol induction in shake flasks. Approximately 1.6 mg of high purity rPCP was obtained from a 100-mL culture by Ni+-affinity chromatography, and sodium dodecyl sulfate polyacrylamide gel electrophoresis results indicated rPCP as a homologous dimer glycoprotein formed by different molecular-weight monomers. Peptide-N-glycosidase F-mediated deglycosylation analysis showed the presence of an N-glycosylated rPCP monomer, and bioactivity assays showed that rPCP activity upregulated tumor necrosis factor (TNF)-α and interleukin-1ß transcription and increased TNF-α secretion from mouse macrophage RAW 264.7 cells. Shortly, we demonstrated successful purification of active rPCP from P. pastoris, which promoted further study of its biological activities and medical applications.


Assuntos
Proteínas Fúngicas/genética , Pichia/genética , Proteínas Recombinantes/genética , Wolfiporia/metabolismo , Animais , Clonagem Molecular , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/farmacologia , Glicosilação , Interleucina-1beta/genética , Camundongos , Pichia/crescimento & desenvolvimento , Pichia/metabolismo , Multimerização Proteica , Células RAW 264.7 , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Transfecção , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima , Wolfiporia/genética
19.
Food Chem ; 276: 383-389, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30409609

RESUMO

Traditionally, the root of Pueraria lobata are widely used as a functional food. It was observed that a 70% ethanol extract showed a dose-dependent inhibition towards mushroom tyrosinase. Among the different isolated compounds, calycosin demonstrated potent inhibitory activity against substrates l-tyrosine and l-DOPA, with IC50 of 1.45 ±â€¯0.03 and 7.02 ±â€¯0.46 µM, respectively. Conversely, formononetin and daidzein exhibit weak inhibition. Moreover, kinetic studies revealed calycosin to be a competitive inhibitor for both substrates. Additionally, molecular docking simulation showed that the hydroxyl groups at C-3' and C-7 positions interacted with the catalytic site and peripheral residues, demonstrating a higher affinity toward mushroom tyrosinase. Accordingly, our results suggest that, rather than a mono-substituted hydroxyl or methoxyl group, the presence of a hydroxyl group at C-3' and a methoxyl group at C-4' position of the isoflavone skeleton plays an essential role in the manifestation of anti-browning activity in food products.


Assuntos
Agaricales/enzimologia , Isoflavonas/farmacologia , Monofenol Mono-Oxigenase/antagonistas & inibidores , Raízes de Plantas/química , Pueraria/química , Agaricales/efeitos dos fármacos , Proteínas Fúngicas/farmacologia , Cinética , Simulação de Acoplamento Molecular
20.
ISME J ; 13(3): 588-602, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30301946

RESUMO

Bacteria are the main nutritional competitors of saprophytic fungi during colonization of their ecological niches. This competition involves the mutual secretion of antimicrobials that kill or inhibit the growth of the competitor. Over the last years it has been demonstrated that fungi respond to the presence of bacteria with changes of their transcriptome, but the significance of these changes with respect to competition for nutrients is not clear as functional proof of the antibacterial activity of the induced gene products is often lacking. Here, we report the genome-wide transcriptional response of the coprophilous mushroom Coprinopsis cinerea to the bacteria Bacillus subtilis and Escherichia coli. The genes induced upon co-cultivation with each bacterium were highly overlapping, suggesting that the fungus uses a similar arsenal of effectors against Gram-positive and -negative bacteria. Intriguingly, the induced genes appeare to encode predominantly secreted peptides and proteins with predicted antibacterial activities, which was validated by comparative proteomics of the C. cinerea secretome. Induced members of two putative antibacterial peptide and protein families in C. cinerea, the cysteine-stabilized αß-defensins (Csαß-defensins) and the GH24-type lysozymes, were purified, and their antibacterial activity was confirmed. These results provide compelling evidence that fungi are able to recognize the presence of bacteria and respond with the expression of an arsenal of secreted antibacterial peptides and proteins.


Assuntos
Agaricales/genética , Antibacterianos/metabolismo , Bacillus subtilis/fisiologia , Escherichia coli/fisiologia , Proteínas Fúngicas/metabolismo , Interações Microbianas , Peptídeos/metabolismo , Agaricales/metabolismo , Antibacterianos/farmacologia , Defensinas/genética , Defensinas/metabolismo , Defensinas/farmacologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/farmacologia , Muramidase/genética , Muramidase/metabolismo , Muramidase/farmacologia , Peptídeos/genética , Peptídeos/farmacologia , Proteômica , Transcriptoma
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