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71.
Yoichiro Harada Reto Buser Elsy M. Ngwa Hiroto Hirayama Markus Aebi Tadashi Suzuki 《The Journal of biological chemistry》2013,288(45):32673-32684
Asparagine (N)-linked glycosylation regulates numerous cellular activities, such as glycoprotein quality control, intracellular trafficking, and cell-cell communications. In eukaryotes, the glycosylation reaction is catalyzed by oligosaccharyltransferase (OST), a multimembrane protein complex that is localized in the endoplasmic reticulum (ER). During N-glycosylation in the ER, the protein-unbound form of oligosaccharides (free oligosaccharides; fOSs), which is structurally related to N-glycan, is released into the ER lumen. However, the enzyme responsible for this process remains unidentified. Here, we demonstrate that eukaryotic OST generates fOSs. Biochemical and genetic analyses using mutant strains of Saccharomyces cerevisiae revealed that the generation of fOSs is tightly correlated with the N-glycosylation activity of OST. Furthermore, we present evidence that the purified OST complex can generate fOSs by hydrolyzing dolichol-linked oligosaccharide, the glycan donor substrate for N-glycosylation. The heterologous expression of a single subunit of OST from the protozoan Leishmania major in S. cerevisiae demonstrated that this enzyme functions both in N-glycosylation and generation of fOSs. This study provides insight into the mechanism of PNGase-independent formation of fOSs. 相似文献
72.
In the current model for Glc3Man9GlcNAc2-P-P-Dol assembly, Man5GlcNAc2-
P-P-Dol, Man-P-Dol, and Glc-P-Dol are synthesized on the cytoplasmic face
of the ER and diffuse transversely to the lumenal leaflet where the
synthesis of the lipid-bound precursor oligosaccharide is completed. To
establish the topological sites of Glc-P-Dol synthesis and the
lipid-mediated glucosyltransfer reactions involved in
Glc3Man9GlcNAc2-P-P-Dol synthesis in ER vesicles from pig brain, the
trypsin-sensitivity of Glc-P-Dol synthase activity and the Glc-P-
Dol:Glc0-2Man9GlcNAc2-P-P-Dol glucosyltransferases (GlcTases) was examined
in sealed microsomal vesicles. Since ER vesicles from brain do not contain
glucose 6-phosphate (Glc 6-P) phosphatase activity, the latency of the
lumenally oriented, processing glucosidase I/II activities was used to
assess the intactness of the vesicle preparations. Comparative enzymatic
studies with sealed ER vesicles from brain and kidney, a tissue that
contains Glc 6-P phosphatase, demonstrate the reliability of using the
processing glucosidase activities as latency markers for topological
studies with microsomal vesicles from non-gluconeogenic tissues lacking Glc
6-P phosphatase. The results obtained from the trypsin-sensitivity assays
with sealed microsomal vesicles from brain are consistent with a
topological model in which Glc-P-Dol is synthesized on the cytoplasmic face
of the ER, and subsequently utilized by the three Glc-P-Dol-mediated
GlcTases after "flip-flopping" to the lumenal monolayer.
相似文献
73.
Julia S Bennett Keith A Jolley PFrederick Sparling Nigel J Saunders CAnthony Hart Ian M Feavers Martin CJ Maiden 《BMC biology》2007,5(1):35
Background
Various typing methods have been developed for Neisseria gonorrhoeae, but none provide the combination of discrimination, reproducibility, portability, and genetic inference that allows the analysis of all aspects of the epidemiology of this pathogen from a single data set. Multilocus sequence typing (MLST) has been used successfully to characterize the related organisms Neisseria meningitidis and Neisseria lactamica. Here, the same seven locus Neisseria scheme was used to characterize a diverse collection of N. gonorrhoeae isolates to investigate whether this method would allow differentiation among isolates, and to distinguish these three species. 相似文献74.
Samuel Wanji Ebanga-Echi Eyong Nicholas Tendongfor Che Ngwa Elive Esuka Arnaud Kengne-Ouafo Fabrice Datchoua-Poutcheu Peter Enyong Adrian Hopkins Charles D. Mackenzie 《PLoS neglected tropical diseases》2015,9(11)
Background
Loiasis, a filarial infection caused by Loa loa usually thought to cause relatively minor morbidity, can cause serious and often fatal reactions in patients carrying very high levels of circulating Loa loa microfilariae (mf) following administration of microfilaricidal drugs. An experimental model of this condition would greatly aid the definition of the optimal management of this important clinical presentation.Methodology/Principle Findings
Fifteen baboons (Papio anubis) were infected with 600 infective larvae (L3) isolated from Chrysops vector flies. Animals were observed for any clinical changes; blood samples were collected every 1–2 months for 22 months, and analysed for parasitological, hematological and biochemical profiles using standard techniques. All animals became patent but remained clinically normal throughout the study. The parasitological pre-patent period was between 4–8 months, with a majority (60%) of animals becoming patent by 5 months post infection (MPI); all animals were patent by 8 MPI. Microfilarial loads increased steadily in all animals and reached a peak at 18 MPI. By 10 MPI >70% of animals had mf >8,000 mf/mL, and at 18 MPI >70% of animals had mf >30,000mf/mL with 50% of these animals with mf >50,000mf/mL. Absolute eosinophil, creatinine, Ca2+ and K+ levels were generally above normal values (NV). Positive associations were seen between microfilariaemia and eosinophilia, Hb, Ca2+, and gamma-GT values, whilst significant negative associations were seen between microfilariaemia and potassium, glucose and mononuclear leukocyte levels.Conclusions
Infection of splenectomised baboons with L. loa can induce levels of circulating microfilariae, and corresponding haematological profiles, which parallel those seen in those humans in danger of the severe post-microfilariacide clinical responses. Utilization of this experimental model could contribute to the improved management of the loiasis related adverse responses in humans. 相似文献75.
Alem Mehari Samina Afreen Julius Ngwa Rosanna Setse Alicia N. Thomas Vishal Poddar Wayne Davis Octavius D. Polk Sheik Hassan Alvin V. Thomas 《PloS one》2015,10(10)
BackgroundObesity prevalence in United States (US) adults exceeds 30% with highest prevalence being among blacks. Obesity is known to have significant effects on respiratory function and obese patients commonly report respiratory complaints requiring pulmonary function tests (PFTs). However, there is no large study showing the relationship between body mass index (BMI) and PFTs in healthy African Americans (AA).ObjectiveTo determine the effect of BMI on PFTs in AA patients who did not have evidence of underlying diseases of the respiratory system.MethodsWe reviewed PFTs of 339 individuals sent for lung function testing who had normal spirometry and lung diffusion capacity for carbon monoxide (DLCO) with wide range of BMI.ResultsFunctional residual capacity (FRC) and expiratory reserve volume (ERV) decreased exponentially with increasing BMI, such that morbid obesity resulted in patients breathing near their residual volume (RV). However, the effects on the extremes of lung volumes, at total lung capacity (TLC) and residual volume (RV) were modest. There was a significant linear inverse relationship between BMI and DLCO, but the group means values remained within the normal ranges even for morbidly obese patients.ConclusionsWe showed that BMI has significant effects on lung function in AA adults and the greatest effects were on FRC and ERV, which occurred at BMI values < 30 kg/m2. These physiological effects of weight gain should be considered when interpreting PFTs and their effects on respiratory symptoms even in the absence of disease and may also exaggerate existing lung diseases. 相似文献
76.
A deterministic ordinary differential equation model for the dynamics of malaria transmission that explicitly integrates the demography and life style of the malaria vector and its interaction with the human population is developed and analyzed. The model is different from standard malaria transmission models in that the vectors involved in disease transmission are those that are questing for human blood. Model results indicate the existence of nontrivial disease free and endemic steady states, which can be driven to instability via a Hopf bifurcation as a parameter is varied in parameter space. Our model therefore captures oscillations that are known to exist in the dynamics of malaria transmission without recourse to external seasonal forcing. Additionally, our model exhibits the phenomenon of backward bifurcation. Two threshold parameters that can be used for purposes of control are identified and studied, and possible reasons why it has been difficult to eradicate malaria are advanced. 相似文献
77.
Background
Many plant species show induced responses that protect them against exogenous attacks. These responses involve the production of many different bioactive compounds. Plant species belonging to the Brassicaceae family produce defensive glucosinolates, which may greatly influence their favorable nutritional properties for humans. Each responding compound may have its own dynamic profile and metabolic relationships with other compounds. The chemical background of the induced response is therefore highly complex and may therefore not reveal all the properties of the response in any single model. 相似文献78.
Wouter Wegdam Perry D Moerland Marrije R Buist Emiel Ver Loren van Themaat Boris Bleijlevens Huub CJ Hoefsloot Chris G de Koster Johannes MFG Aerts 《Proteome science》2009,7(1):19-17
Background
Mass spectrometry is increasingly being used to discover proteins or protein profiles associated with disease. Experimental design of mass-spectrometry studies has come under close scrutiny and the importance of strict protocols for sample collection is now understood. However, the question of how best to process the large quantities of data generated is still unanswered. Main challenges for the analysis are the choice of proper pre-processing and classification methods. While these two issues have been investigated in isolation, we propose to use the classification of patient samples as a clinically relevant benchmark for the evaluation of pre-processing methods. 相似文献79.
Matthias Erb Ruth Gordon-Weeks Victor Flors Gemma Cama?es Ted CJ Turlings Jurriaan Ton 《Plant signaling & behavior》2009,4(7):636-638
Plants are important mediators between above- and belowground herbivores. Consequently, interactions between root and shoot defenses can have far-reaching impacts on entire food webs. We recently reported that infestation of maize roots by larvae of the beetle Diabrotica virgifera virgifera induced shoot resistance against herbivores and pathogens. Root herbivory also enhanced aboveground DIMBOA and primed for enhanced induction of chlorogenic acid, two secondary metabolites that have been associated with plant stress resistance. Interestingly, the plant hormone abscisic acid (ABA) emerged as a putative long-distance signal in the regulation of these systemic defenses. In this addendum, we have investigated the role of root-derived ABA in aboveground regulation of DIMBOA and the phenolic compounds chlorogenic acid, caffeic and ferulic acid. Furthermore, we discuss the relevance of ABA in relation to defense against the leaf herbivore Spodoptera littoralis. Soil-drench treatment with ABA mimicked root herbivore-induced accumulation of DIMBOA in the leaves. Similarly, ABA mimicked aboveground priming of chlorogenic acid production, causing augmented induction of this compound after subsequent shoot attack by S. littoralis caterpillars. These findings confirm our notion that ABA acts as an important signal in the regulation of aboveground defenses during belowground herbivory. However, based on our previous finding that ABA alone is not sufficient to trigger aboveground resistance against S. littoralis caterpillars, our results also suggest that the ABA-inducible effects on DIMBOA and chlorogenic acid are not solely responsible for root herbivore-induced resistance against S. littoralis.Key words: induced resistance, Spodoptera littoralis, Zea mays, Diabrotica virgifera, DIMBOA, chlorogenic acid, absisic acid, priming 相似文献
80.