Base de dados : MEDLINE
Pesquisa : B01.300.179.100.125 [Categoria DeCS]
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[PMID]:28825818
[Au] Autor:Reddy MD; Kobori H; Mori T; Wu J; Kawagishi H; Watkins EB
[Ad] Endereço:Department of Pharmaceutical Sciences, College of Pharmacy, Union University , Jackson, Tennessee 38305, United States.
[Ti] Título:Gram-Scale, Stereoselective Synthesis and Biological Evaluation of (+)-Armillariol C.
[So] Source:J Nat Prod;80(9):2561-2565, 2017 Sep 22.
[Is] ISSN:1520-6025
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Natural products with heteroaromatic cores are ample and widespread in nature, with many compounds exhibiting promising therapeutic properties. (+)-Armillariol C (1a) is a furan-based natural product isolated from Armillaria species. Herein, we report the first enantioselective synthesis of (+)-armillariol C (1a, 79% overall yield), its enantiomer (1b), and four other analogues, on a gram-scale, using microwave-mediated, Suzuki-Miyaura cross-coupling and Sharpless asymmetric dihydroxylation reactions. Compounds were tested for plant- and mycelia-growth regulatory activity, with 1b, 7a, and 7b showing the strongest inhibitory properties in a lettuce assay and 7b and 9b inhibiting Flammulina velutipes.
[Mh] Termos MeSH primário: Armillaria/química
Produtos Biológicos/síntese química
Furanos/síntese química
[Mh] Termos MeSH secundário: Evolução Biológica
Produtos Biológicos/química
Produtos Biológicos/isolamento & purificação
Furanos/química
Furanos/isolamento & purificação
Estrutura Molecular
Estereoisomerismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biological Products); 0 (Furans); 0 (armillariol C)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171031
[Lr] Data última revisão:
171031
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170822
[St] Status:MEDLINE
[do] DOI:10.1021/acs.jnatprod.7b00484


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[PMID]:28726108
[Au] Autor:Purtov KV; Petushkov VN; Rodionova NS; Gitelson JI
[Ad] Endereço:Institute of Biophysics, Krasnoyarsk Research Center, Siberian Branch, Russian Academy of Sciences, Akademgorodok, Krasnoyarsk, 660036, Russia. purtovk@mail.ru.
[Ti] Título:Why does the bioluminescent fungus Armillaria mellea have luminous mycelium but nonluminous fruiting body?
[So] Source:Dokl Biochem Biophys;474(1):217-219, 2017 May.
[Is] ISSN:1608-3091
[Cp] País de publicação:Russia (Federation)
[La] Idioma:eng
[Ab] Resumo:By determining the components involved in the bioluminescence process in luminous and nonluminous organs of the honey fungus Armillaria mellea, we have established causes of partial luminescence of this fungus. The complete set of enzymes and substrates required for bioluminescence is formed only in the mycelium and only under the conditions of free oxygen access. Since the synthesis of luciferin precursor (hispidin) and 3-hydroxyhispidin hydroxylase in the fruiting bodies is blocked, the formation of luciferin-the key component of fungal bioluminescent system-was not observed. That is why the fruiting body of Armillaria mellea is nonluminous despite the presence of luciferase, the enzyme that catalyzes the oxidation of luciferin with a photon emission.
[Mh] Termos MeSH primário: Armillaria/metabolismo
Carpóforos/metabolismo
Luminescência
Micélio/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170906
[Lr] Data última revisão:
170906
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170721
[St] Status:MEDLINE
[do] DOI:10.1134/S1607672917030176


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[PMID]:28284022
[Au] Autor:Lukanc T; Brzin J; Kos J; Sabotic J
[Ad] Endereço:Department of Biotechnology, Jozef Stefan Institute, SI1000 Ljubljana, Slovenia.
[Ti] Título:Trypsin-specific Inhibitors from the Macrolepiota procera, Armillaria mellea and Amanita phalloides wild mushrooms.
[So] Source:Acta Biochim Pol;64(1):21-24, 2017.
[Is] ISSN:1734-154X
[Cp] País de publicação:Poland
[La] Idioma:eng
[Ab] Resumo:Wild growing mushrooms are a rich source of novel proteins with unique features. We have isolated and characterized trypsin inhibitors from two edible mushrooms, the honey fungus (Armillaria mellea) and the parasol mushroom (Macrolepiota procera), and from the poisonous death cap (Amanita phalloides). The trypsin inhibitors isolated: armespin, macrospin and amphaspin, have similar molecular masses, acidic isoelectric points and are not N-glycosylated. They are very strong trypsin inhibitors and weak chymotrypsin inhibitors. They are resistant to exposure to high temperatures and withstand extreme pH values. These exceptional characteristics are advantageous for their potential use in biotechnology, agriculture and medicine.
[Mh] Termos MeSH primário: Amanita/química
Armillaria/química
Inibidores da Tripsina/isolamento & purificação
[Mh] Termos MeSH secundário: Temperatura Alta
Concentração de Íons de Hidrogênio
Ponto Isoelétrico
Peso Molecular
Inibidores da Tripsina/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Trypsin Inhibitors)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170417
[Lr] Data última revisão:
170417
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170312
[St] Status:MEDLINE
[do] DOI:10.18388/abp.2015_1187


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[PMID]:28257665
[Au] Autor:Geng Y; Zhu S; Cheng P; Lu ZM; Xu HY; Shi JS; Xu ZH
[Ad] Endereço:School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
[Ti] Título:Bioassay-guided fractionation of ethyl acetate extract from Armillaria mellea attenuates inflammatory response in lipopolysaccharide (LPS) stimulated BV-2 microglia.
[So] Source:Phytomedicine;26:55-61, 2017 Mar 15.
[Is] ISSN:1618-095X
[Cp] País de publicação:Germany
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Armillaria mellea (A. mellea) is a traditional Chinese medicinal and edible mushroom, which is proved to possess a lot of biological activities, including anti-oxidation, immunopotentiation, anti-vertigo and anti-aging activities. However, little information is available in regard to its neuroprotection activity in inflammation-mediated neurodegenerative diseases. PURPOSE: We have found that A. mellea has an anti-inflammatory activity in LPS-induced RAW264.7 cells in our previous study. The objective of this study is to investigate the anti-neuroinflammatory mechanism of a bioassay-guided fractionation (Fr.2) and its active components/compounds. METHODS: Compounds were isolated by preparative high performance liquid chromatography (pre-HPLC) and their structures were established by mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopic analyses. The anti-neuroinflammatory effect of Fr.2 and each compounds were investigated in lipopolysaccharide (LPS)-stimulated murine microglia cell lineBV-2. RESULTS: We demonstrated that Fr.2 significantly decreased the production of inflammation mediator nitric oxide (NO) and inflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1beta (IL-1ß) in a dose-dependent manner (10, 30, 100µg/ml). In addition, Fr.2 markedly down-regulated the phosphorylation levels of nuclear factor kappa B p65 (NF-κB p65), inhibitory κB-α (IκB-α) and c-Jun N-terminal kinases (JNKs) pathways. Sevens compounds were isolated from Fr.2, among them, three compounds, 5-hydroxymethylfurfural (CP1), vanillic acid (CP4) and syringate (CP5) were reported for the first time in A. mellea. NO and inflammatory cytokines (TNF-α, IL-6, IL-1ß) secretion indicated that daidzein (CP6) and genistein (CP7) showed a more outstanding anti-inflammation potential at non-toxic concentrations (10, 30, 100µM) than the other five compounds. CONCLUSIONS: In conclusion, Fr.2 may have therapeutic potential for neurodegenerative diseases by inhibiting inflammatory mediators and suppress inflammation pathway in activated microglia. Daidzein and genistein may serve as the effective anti-inflammation compounds of Fr.2.
[Mh] Termos MeSH primário: Acetatos/farmacologia
Anti-Inflamatórios/farmacologia
Inflamação/tratamento farmacológico
Lipopolissacarídeos/efeitos adversos
Microglia/efeitos dos fármacos
Fármacos Neuroprotetores/farmacologia
Extratos Vegetais/farmacologia
[Mh] Termos MeSH secundário: Animais
Armillaria/química
Medicamentos de Ervas Chinesas/farmacologia
Camundongos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Acetates); 0 (Anti-Inflammatory Agents); 0 (Drugs, Chinese Herbal); 0 (Lipopolysaccharides); 0 (Neuroprotective Agents); 0 (Plant Extracts); 76845O8NMZ (ethyl acetate)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170517
[Lr] Data última revisão:
170517
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170305
[St] Status:MEDLINE


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[PMID]:28183153
[Au] Autor:Chen WB; Cheng MJ; Tian YB; Wang QH; Wang B; Li MJ; Fang RJ
[Ad] Endereço:College of Animal Science and Technology, Hunan Agricultural University, Changsha, China.
[Ti] Título:Effects of Armillariella tabescens mycelia on the growth performance and intestinal immune response and microflora of early-weaned pigs.
[So] Source:Anim Sci J;88(9):1388-1397, 2017 Sep.
[Is] ISSN:1740-0929
[Cp] País de publicação:Australia
[La] Idioma:eng
[Ab] Resumo:This study was performed to evaluate effects of Armillariella tabescens (A. tabescens) on the growth performance and intestinal immune response and microflora in early-weaned pigs when used as feed additive. A. tabescens mycelia were added to basal diets at concentrations of 0%, 0.1%, 0.3% or 0.9% (w/w). A total of 144 commercial cross-bred piglets were randomly allocated to one of these four diets and fed for 30 days. The growth performance of early-weaned piglets displayed improvement with diets containing 0.1% and 0.3% dried mycelia powder from A. tabescens. Supplementing with 0.1% or 0.3% A. tabescens mycelia induced a 2.6- and three-fold increase in secretory immunoglobulin A (sIgA) content in the jejunal mucosa, respectively, but had only a marginal effect on sIgA in the ileal mucosa. Expression of interleukin-2, interferon-γ, and tumor necrosis factor-α in the jejunal mucosa were elevated with A. tabescens mycelia administration. Increased amounts of Lactobacillus spp. and Bifidobacterium spp. in the jejunum, and decreased amounts of Escherichia coli in the jejunum and ileum were observed with the administration of A. tabescens-containing diets. This study demonstrated that A. tabescens had beneficial effects on the growth performance and intestinal microflora of early-weaned pigs.
[Mh] Termos MeSH primário: Ração Animal
Armillaria
Dieta/veterinária
Suplementos Nutricionais
Microbioma Gastrointestinal
Intestinos/imunologia
Intestinos/microbiologia
Suínos/crescimento & desenvolvimento
Suínos/imunologia
Suínos/microbiologia
[Mh] Termos MeSH secundário: Animais
Feminino
Íleo/metabolismo
Imunoglobulina A Secretora/metabolismo
Interferon gama/metabolismo
Interleucina-2/metabolismo
Mucosa Intestinal/metabolismo
Jejuno/metabolismo
Masculino
Pós
Fator de Necrose Tumoral alfa/metabolismo
Desmame
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Immunoglobulin A, Secretory); 0 (Interleukin-2); 0 (Powders); 0 (Tumor Necrosis Factor-alpha); 82115-62-6 (Interferon-gamma)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171005
[Lr] Data última revisão:
171005
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170210
[St] Status:MEDLINE
[do] DOI:10.1111/asj.12765


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[PMID]:28122504
[Au] Autor:Koch RA; Wilson AW; Séné O; Henkel TW; Aime MC
[Ad] Endereço:Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN, 47907, USA.
[Ti] Título:Resolved phylogeny and biogeography of the root pathogen Armillaria and its gasteroid relative, Guyanagaster.
[So] Source:BMC Evol Biol;17(1):33, 2017 Jan 25.
[Is] ISSN:1471-2148
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Armillaria is a globally distributed mushroom-forming genus composed primarily of plant pathogens. Species in this genus are prolific producers of rhizomorphs, or vegetative structures, which, when found, are often associated with infection. Because of their importance as plant pathogens, understanding the evolutionary origins of this genus and how it gained a worldwide distribution is of interest. The first gasteroid fungus with close affinities to Armillaria-Guyanagaster necrorhizus-was described from the Neotropical rainforests of Guyana. In this study, we conducted phylogenetic analyses to fully resolve the relationship of G. necrorhizus with Armillaria. Data sets containing Guyanagaster from two collecting localities, along with a global sampling of 21 Armillaria species-including newly collected specimens from Guyana and Africa-at six loci (28S, EF1α, RPB2, TUB, actin-1 and gpd) were used. Three loci-28S, EF1α and RPB2-were analyzed in a partitioned nucleotide data set to infer divergence dates and ancestral range estimations for well-supported, monophyletic lineages. RESULTS: The six-locus phylogenetic analysis resolves Guyanagaster as the earliest diverging lineage in the armillarioid clade. The next lineage to diverge is that composed of species in Armillaria subgenus Desarmillaria. This subgenus is elevated to genus level to accommodate the exannulate mushroom-forming armillarioid species. The final lineage to diverge is that composed of annulate mushroom-forming armillarioid species, in what is now Armillaria sensu stricto. The molecular clock analysis and ancestral range estimation suggest the most recent common ancestor to the armillarioid lineage arose 51 million years ago in Eurasia. A new species, Guyanagaster lucianii sp. nov. from Guyana, is described. CONCLUSIONS: The armillarioid lineage evolved in Eurasia during the height of tropical rainforest expansion about 51 million years ago, a time marked by a warm and wet global climate. Species of Guyanagaster and Desarmillaria represent extant taxa of these early diverging lineages. Desarmillaria represents an armillarioid lineage that was likely much more widespread in the past. Guyanagaster likely evolved from a gilled mushroom ancestor and could represent a highly specialized endemic in the Guiana Shield. Armillaria species represent those that evolved after the shift in climate from warm and tropical to cool and arid during the late Eocene. No species in either Desarmillaria or Guyanagaster are known to produce melanized rhizomorphs in nature, whereas almost all Armillaria species are known to produce them. The production of rhizomorphs is an adaptation to harsh environments, and could be a driver of diversification in Armillaria by conferring a competitive advantage to the species that produce them.
[Mh] Termos MeSH primário: Armillaria/classificação
Basidiomycota/classificação
Raízes de Plantas/microbiologia
[Mh] Termos MeSH secundário: África
Clima
Evolução Molecular
Guiana
Filogenia
Filogeografia
Análise de Sequência de DNA
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170808
[Lr] Data última revisão:
170808
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170127
[St] Status:MEDLINE
[do] DOI:10.1186/s12862-017-0877-3


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[PMID]:27138686
[Au] Autor:Guo T; Wang HC; Xue WQ; Zhao J; Yang ZL
[Ad] Endereço:Key Laboratory for Plant Diversity and Biogeography of East Asia, Kunming Institute of Botany, Chinese Academy of Sciences, Heilongtan, Kunming 650201, China.
[Ti] Título:Phylogenetic Analyses of Armillaria Reveal at Least 15 Phylogenetic Lineages in China, Seven of Which Are Associated with Cultivated Gastrodia elata.
[So] Source:PLoS One;11(5):e0154794, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Fungal species of Armillaria, which can act as plant pathogens and/or symbionts of the Chinese traditional medicinal herb Gastrodia elata ("Tianma"), are ecologically and economically important and have consequently attracted the attention of mycologists. However, their taxonomy has been highly dependent on morphological characterization and mating tests. In this study, we phylogenetically analyzed Chinese Armillaria samples using the sequences of the internal transcribed spacer region, translation elongation factor-1 alpha gene and beta-tubulin gene. Our data revealed at least 15 phylogenetic lineages of Armillaria from China, of which seven were newly discovered and two were recorded from China for the first time. Fourteen Chinese biological species of Armillaria, which were previously defined based on mating tests, could be assigned to the 15 phylogenetic lineages identified herein. Seven of the 15 phylogenetic lineages were found to be disjunctively distributed in different continents of the Northern Hemisphere, while eight were revealed to be endemic to certain continents. In addition, we found that seven phylogenetic lineages of Armillaria were used for the cultivation of Tianma, only two of which had been recorded to be associated with Tianma previously. We also illustrated that G. elata f. glauca ("Brown Tianma") and G. elata f. elata ("Red Tianma"), two cultivars of Tianma grown in different regions of China, form symbiotic relationships with different phylogenetic lineages of Armillaria. These findings should aid the development of Tianma cultivation in China.
[Mh] Termos MeSH primário: Armillaria/genética
Armillaria/fisiologia
Gastrodia/microbiologia
Filogenia
[Mh] Termos MeSH secundário: China
Evolução Molecular
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170713
[Lr] Data última revisão:
170713
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160504
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0154794


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[PMID]:26952552
[Au] Autor:Li Z; Wang Y; Jiang B; Li W; Zheng L; Yang X; Bao Y; Sun L; Huang Y; Li Y
[Ad] Endereço:National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun 130024, China.
[Ti] Título:Structure, cytotoxic activity and mechanism of protoilludane sesquiterpene aryl esters from the mycelium of Armillaria mellea.
[So] Source:J Ethnopharmacol;184:119-27, 2016 May 26.
[Is] ISSN:1872-7573
[Cp] País de publicação:Ireland
[La] Idioma:eng
[Ab] Resumo:ETHNOPHARMACOLOGICAL RELEVANCE: Armillaria mellea (Vahl. ex. Fr.) Karst is an important traditional Chinese medicine used in dispelling wind and removing obstruction in the meridians, and strengthening tendons and bones. Armillaria mellea has been recorded in the book Caobenshiyi which was written by ancestor for the function of suppressing hyderactive liver for calming endogenous wind medicine. The aim of this study is to investigate the cytotoxic activity for liver cell lines (normal and cancerous) of protoilludane sesquiterpene aryl esters from the mycelium of A. mellea. MATERIALS AND METHODS: A systemic fractionation of the mycelium extracts of A. mellea and relative activity mechanisms were studied. RESULTS: Two new protoilludane sesquiterpene aryl esters named 5'-methoxy-armillasin (1) and 5-hydroxyl-armillarivin (2) were isolated. In addition, eight known protoilludane sesquiterpene aryl esters armillaridin (3), armillartin (4), armillarin (5), melleolide B (6), armillarilin (7), armillasin (8), armillarigin (9) and melleolide (10) were also isolated from the mycelium of A. mellea. The relative configurations of the two new compounds were confirmed by NOESY spectra. Among ten protoilludane sesquiterpene aryl esters, compounds 2, 3, 4, 7, 8, 9 and 10 were active constituents with highly cytotoxic activity against HepG2 cells (4.95-37.65µg/mL). We reported here for the time, that compound 10 (melleolide) showed anti-tumor ability on hepatoma cell. The relative mechanism was assessed on HepG2 cells. CONCLUSIONS: Among all the ten protoilludane sesquiterpene aryl esters, melleolide (10) showed the best cytotoxic activity for HepG2 cells (4.95µg/mL) and lower activity for L02 cells (16.05µg/mL). Mechanism study showed that melleolide decreased the viability of the cancer cells with varying levels of cleaved-caspase 3, caspase 8, caspase 9, Bax and Ki67 expression. On the other hand, melleolide induced HepG2 cell cycle arrest at the G2/M phase.
[Mh] Termos MeSH primário: Antineoplásicos/farmacologia
Armillaria
Sesquiterpenos/farmacologia
[Mh] Termos MeSH secundário: Antineoplásicos/química
Antineoplásicos/isolamento & purificação
Apoptose/efeitos dos fármacos
Ciclo Celular/efeitos dos fármacos
Linhagem Celular
Linhagem Celular Tumoral
Sobrevivência Celular/efeitos dos fármacos
Ésteres
Seres Humanos
Estrutura Molecular
Micélio/química
Sesquiterpenos/química
Sesquiterpenos/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antineoplastic Agents); 0 (Esters); 0 (Sesquiterpenes)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170106
[Lr] Data última revisão:
170106
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160309
[St] Status:MEDLINE


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[PMID]:26845767
[Au] Autor:Ødum AS; Østergaard S; Nørby I; Meldal M; Olesen K
[Ti] Título:Novel Application of Peptidyl-Lys Metallopeptidase as a C-Terminal Processing Protease.
[So] Source:Protein Pept Lett;23(4):396-403, 2016.
[Is] ISSN:1875-5305
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Adding fusion partners to proteins or peptides can aid or be a necessity to facilitate recombinant expression, folding, or purification. Independent of the reason it is desirable to remove the fusion partner to restore native functionality. Processing proteases catalyze the removal of fusion partners, however, most of these proteases have substrate specificity for the N-terminal of the scissile bond, leaving non-native termini if fusions are added to the C-terminal. The peptidyl-lys metallopeptidease of Armillaria mellea (Am-LysN) is unusual by having substrate specificity for the C-terminal side of the scissile peptide bond, allowing it to generate native C-termini. Am-LysN has strict specificity for lysine in P1', making all lysines of a protein or peptide a potential degradation site, however there are a number of amino acid side chains which lower hydrolysis significantly when located adjacent to the lysine. In this study we show that Am-LysN can be used as a processing protease to remove C-terminal extensions of peptides with no internal lysine to generate native Ctermini. Furthermore we show that removal of C-terminal extensions on peptides containing internal lysines can be achieved with little degradation of the product depending on the adjacent amino acids. These results demonstrate the utility of LysN allowing for novel ways to use fusion technology in the production of recombinant proteins.
[Mh] Termos MeSH primário: Armillaria/enzimologia
Metaloproteases/química
Peptídeos/química
[Mh] Termos MeSH secundário: Armillaria/química
Lisina/metabolismo
Conformação Proteica
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Peptides); EC 3.4.- (AMMP protein, Armillaria mellea); EC 3.4.- (Metalloproteases); K3Z4F929H6 (Lysine)
[Em] Mês de entrada:1612
[Cu] Atualização por classe:161230
[Lr] Data última revisão:
161230
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160205
[St] Status:MEDLINE


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[PMID]:26572161
[Au] Autor:Ødum AS; Østergaard S; Nørby I; Meldal M; Olesen K
[Ad] Endereço:Global Research, Novo Nordisk A/S, Novo Nordisk Park, 2760 Måløv, Denmark and Center for Evolutionary Chemical Biology, Nano-Science Center, University of Copenhagen, Universitetsparken 5, 2100 København Ø, Denmark.
[Ti] Título:Heterologous expression of peptidyl-Lys metallopeptidase of Armillaria mellea and mutagenic analysis of the recombinant peptidase.
[So] Source:J Biochem;159(4):461-70, 2016 Apr.
[Is] ISSN:1756-2651
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A method to express, purify and modify the Peptidyl-Lys metallopeptidase (LysN) ofArmillaria melleainPichia pastoriswas developed to enable functional studies of the protease. Based on prior work, we propose a mechanism of action of LysN. Catalytic residues were investigated by site-directed mutagenesis. As anticipated, these mutations resulted in significantly reduced catalytic rates. Additionally, based on molecular modelling eleven mutants were designed to have altered substrate specificity. The S1' binding pocket of LysN is quite narrow and lined with negative charge to specifically accommodate lysine. To allow for arginine specificity in S1', it was proposed to extend the S1' binding pocket by mutagenesis, however the resulting mutant did not show any activity with arginine in P1'. Two mutants, A101D and T105D, showed increased specificity towards arginine in subsites S2'-S4' compared to the wild type protease. We speculate that the increased specificity to result from the additional negative charge which attract and interact with positively charged residues better than the wild type.
[Mh] Termos MeSH primário: Arginina/química
Armillaria/enzimologia
Proteínas Fúngicas/química
Metaloendopeptidases/química
Proteínas Recombinantes/química
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Sítios de Ligação
Catálise
Ensaios Enzimáticos
Proteínas Fúngicas/genética
Proteínas Fúngicas/isolamento & purificação
Cinética
Metaloendopeptidases/genética
Metaloendopeptidases/isolamento & purificação
Modelos Moleculares
Dados de Sequência Molecular
Mutagênese Sítio-Dirigida
Pichia/genética
Proteínas Recombinantes/genética
Proteínas Recombinantes/isolamento & purificação
Homologia de Sequência de Aminoácidos
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Fungal Proteins); 0 (Recombinant Proteins); 94ZLA3W45F (Arginine); EC 3.4.24.- (Metalloendopeptidases); EC 3.4.24.20 (peptidyl-Lys metalloendopeptidase)
[Em] Mês de entrada:1610
[Cu] Atualização por classe:170403
[Lr] Data última revisão:
170403
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:151118
[St] Status:MEDLINE
[do] DOI:10.1093/jb/mvv115



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