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41.
Jacob Ball Renata A. G. Reis Johnson Agniswamy Irene T. Weber Giovanni Gadda 《Protein science : a publication of the Protein Society》2019,28(1):167-175
The crystal structure of the NADH:quinone oxidoreductase PA1024 has been solved in complex with NAD+ to 2.2 Å resolution. The nicotinamide C4 is 3.6 Å from the FMN N5 atom, with a suitable orientation for facile hydride transfer. NAD+ binds in a folded conformation at the interface of the TIM‐barrel domain and the extended domain of the enzyme. Comparison of the enzyme‐NAD+ structure with that of the ligand‐free enzyme revealed a different conformation of a short loop (75–86) that is part of the NAD+‐binding pocket. P78, P82, and P84 provide internal rigidity to the loop, whereas Q80 serves as an active site latch that secures the NAD+ within the binding pocket. An interrupted helix consisting of two α‐helices connected by a small three‐residue loop binds the pyrophosphate moiety of NAD+. The adenine moiety of NAD+ appears to π–π stack with Y261. Steric constraints between the adenosine ribose of NAD+, P78, and Q80, control the strict specificity of the enzyme for NADH. Charged residues do not play a role in the specificity of PA1024 for the NADH substrate. 相似文献
42.
Diego de Souza Gonalves Marina da Silva Ferreira Kamilla Xavier Gomes Claudia Rodríguez‐de La Noval Susie Coutinho Liedke Giovani Carlo Veríssimo da Costa Patricia Albuquerque Juliana Reis Cortines Regina Helena Saramago Peralta Jos Mauro Peralta Arturo Casadevall Allan J. Guimares 《Cellular microbiology》2019,21(10)
Free‐living amoebae (FLAs) are major reservoirs for a variety of bacteria, viruses, and fungi. The most studied mycophagic FLA, Acanthamoeba castellanii (Ac), is a potential environmental host for endemic fungal pathogens such as Cryptococcus spp., Histoplasma capsulatum, Blastomyces dermatitides, and Sporothrix schenckii. However, the mechanisms involved in this interaction are poorly understood. The aim of this work was to characterize the molecular instances that enable Ac to interact with and ingest fungal pathogens, a process that could lead to selection and maintenance of possible virulence factors. The interaction of Ac with a variety of fungal pathogens was analysed in a multifactorial evaluation that included the role of multiplicity of infection over time. Fungal binding to Ac surface by living image consisted of a quick process, and fungal initial extrusion (vomocytosis) was detected from 15 to 80 min depending on the organism. When these fungi were cocultured with the amoeba, only Candida albicans and Cryptococcus neoformans were able to grow, whereas Paracoccidioides brasiliensis and Sporothrix brasiliensis displayed unchanged viability. Yeasts of H. capsulatum and Saccharomyces cerevisiae were rapidly killed by Ac; however, some cells remained viable after 48 hr. To evaluate changes in fungal virulence upon cocultivation with Ac, recovered yeasts were used to infect Galleria mellonella, and in all instances, they killed the larvae faster than control yeasts. Surface biotinylated extracts of Ac exhibited intense fungal binding by FACS and fluorescence microscopy. Binding was also intense to mannose, and mass spectrometry identified Ac proteins with affinity to fungal surfaces including two putative transmembrane mannose‐binding proteins (MBP, L8WXW7 and MBP1, Q6J288). Consistent with interactions with such mannose‐binding proteins, Ac–fungi interactions were inhibited by mannose. These MBPs may be involved in fungal recognition by amoeba and promotes interactions that allow the emergence and maintenance of fungal virulence for animals. 相似文献
43.
Bingxin Wang Daniel G. Rocha Mark I. Abrahams Andr P. Antunes Hugo C. M. Costa Andr Luis Sousa Gonalves Wilson Roberto Spironello Milton Jos de Paula Carlos A. Peres Juarez Pezzuti Emiliano Ramalho Marcelo Lima Reis Elildo Carvalho Jr Fabio Rohe David W. Macdonald Cedric Kai Wei Tan 《Ecology and evolution》2019,9(9):5049-5062
Amazonia forest plays a major role in providing ecosystem services for human and sanctuaries for wildlife. However, ongoing deforestation and habitat fragmentation in the Brazilian Amazon has threatened both. The ocelot is an ecologically important mesopredator and a potential conservation ambassador species, yet there are no previous studies on its habitat preference and spatial patterns in this biome. From 2010 to 2017, twelve sites were surveyed, totaling 899 camera trap stations, the largest known dataset for this species. Using occupancy modeling incorporating spatial autocorrelation, we assessed habitat use for ocelot populations across the Brazilian Amazon. Our results revealed a positive sigmoidal correlation between remote‐sensing derived metrics of forest cover, disjunct core area density, elevation, distance to roads, distance to settlements and habitat use, and that habitat use by ocelots was negatively associated with slope and distance to river/lake. These findings shed light on the regional scale habitat use of ocelots and indicate important species–habitat relationships, thus providing valuable information for conservation management and land‐use planning. 相似文献
44.
Leaf Decomposition in a Dry Season Irrigation Experiment in Eastern Amazonian Forest Regrowth 总被引:1,自引:0,他引:1
Steel Silva Vasconcelos Daniel Jacob Zarin Maria Beatriz Silva da Rosa Francisco de Assis Oliveira Cláudio José Reis de Carvalho 《Biotropica》2007,39(5):593-600
Leaf-litter decomposition is a major component of carbon and nutrient dynamics in tropical forest ecosystems, and moisture availability is widely considered to be a major influence on decomposition rates. Here, we report the results of a study of leaf-litter decomposition of five tree species in response to dry-season irrigation in a tropical forest regrowth stand in the Brazilian Amazon; three experiments differing in the timing of installation and duration allowed for an improved resolution of irrigation effects on decomposition. We hypothesized that decomposition rates would be faster under higher moisture availability in the wet season and during dry-season irrigation periods in the treatment plots, and that decomposition rates would be faster for species with higher quality leaves, independent of treatment. The rates of decomposition ( k ) were up to 2.4 times higher in irrigated plots than in control plots. The highest k values were shown by Annona paludosa (0.97 to 1.26/yr) while Ocotea guianensis (0.73 to 0.85/yr) had the lowest values; intermediate rates were found for Lacistema pubescens (0.91 to 1.02/yr) and Vismia guianensis (0.91 to 1.08/yr). These four tree species differed significantly in leaf-litter quality parameters (nitrogen, phosphorus, lignin, and cellulose concentrations, as well as lignin:nitrogen and carbon:nitrogen ratios), but differences in decomposition rates among tree species were not strictly correlated with leaf-litter quality. Overall, our results show that dry-season moisture deficits limit decomposition in Amazonian forest regrowth. 相似文献
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There is strong interest in creating an enzyme that can deacylate natural cephalosporins such as cephalosporin C in order to efficiently acquire the starting compound for the industrial production of semisynthetic cephalosporin antibiotics. In this study, the active site of the glutaryl acylase from Pseudomonas SY-77 was randomized rationally. Several mutations that were found in previous studies to enhance the activity of the enzyme towards adipyl-7-aminodesacetoxycephalosporanic acid (ADCA) and cephalosporin C have now been combined, and libraries have been made in which random amino acid substitutions at these positions are joined. The mutants were expressed in a leucine-deficient Escherichia coli strain and subjected to growth selection with adipyl-leucine or amino-adipyl-leucine as sole leucine source. The mutants growing on these media were selected and purified, and their hydrolysis activities towards adipyl-7-ADCA and cephalosporin C were tested. Several mutants with highly improved activities towards the desired substrates were found in these rationally randomized libraries. The best mutant was selected from a library of totally randomized residues: 178, 266, and 375. This mutant comprises two mutations, Y178F + F375H, which synergistically improve the catalytic efficiency towards adipyl-7-ADCA 36-fold. The activity of this mutant towards adipyl-7-ADCA is 50% of the activity of the wild-type enzyme towards the preferred substrate glutaryl-7-aminocephalosporanic acid, and therefore the characteristics of this mutant approach those needed for industrial application. 相似文献
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Alirya Magda Santos do Vale Gomes Fabrício de Oliveira Reis Raimunda Nonata Santos de Lemos Janaína Marques Mondego Heder Braun José Ribamar Gusmão Araujo 《Revista Brasileira de Entomologia》2019,63(2):119-123
Aleurocanthus woglumi (Ashby, 1915) is an important agricultural pest that causes yield losses of 20–80% in citrus plants by removing plant nutrients while feeding and allowing the formation of sooty mold. The objective of this study was to evaluate physiological changes in citrus plants in response to A. woglumi infestation under field conditions. The experiment was conducted in a citrus orchard in Paço do Lumiar, Maranhão, Brazil. Thirty-two citrus plants were used, including eight of each of the following varieties: Tahiti lime, Tanjaroa tangerine, Nissey tangerine, and Ponkan tangerine. Four random plants with A. woglumi infestation and four plants free from this pest were selected from each variety. The physiological parameters evaluated were photochemical efficiency and gas exchange. Regarding photochemical efficiency, infested plants presented photoinhibition damage, with a performance index of 4.22. The gas exchange parameters of infested plants changed, with reductions in photosynthetic CO2 assimilation of 69.7% (Tahiti), 64% (Tanjaroa), 68.8% (Nissey) and 63.3% (Ponkan). Plants infested with A. woglumi also presented physiological changes; their photosynthetic CO2 assimilation, stomatal conductance, instantaneous transpiration, and performance indexes were affected. The infested citrus plants showed photoinhibition of photosystem II. The photosynthetic CO2 assimilation decreased approximately 70% in Tahiti lime, Tanjaroa tangerine, Nissey tangerine, and Ponkan tangerine plants infested with A. woglumi. 相似文献
50.