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91.
Sabbía V Piovani R Naya H Rodríguez-Maseda H Romero H Musto H 《Journal of biomolecular structure & dynamics》2007,25(1):55-59
Correspondence analysis of amino acid usage was applied to 14,815 complete proteins from the human genome. We found that three major factors influence the variability of amino acidic composition of these proteins, explaining, respectively 20.4%, 14.7%, and 9.9% of the total variability. The first trend is strongly correlated with the GC content of first and second codon positions and is also significantly correlated with the GC level of the corresponding flanking regions and introns. Therefore, the main force shaping amino acid usage among human proteins are the compositional constraints determined by the isochore in which each gene is embedded. The second trend correlates with the hydropathy of each protein and with the frequency of beta-strands. Finally, the third trend is strongly associated with the usage of Cys and the frequency of alpha-helices. 相似文献
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Daniele P. Romancino Valentina Buffa Stefano Caruso Ines Ferrara Samuele Raccosta Antonietta Notaro Yvan Campos Rosina Noto Vincenzo Martorana Antonio Cupane Agata Giallongo Alessandra dAzzo Mauro Manno Antonella Bongiovanni 《Biochimica et Biophysica Acta (BBA)/General Subjects》2018,1862(12):2879-2887
Background
Virtually all cell types have the capacity to secrete nanometer-sized extracellular vesicles, which have emerged in recent years as potent signal transducers and cell-cell communicators. The multifunctional protein Alix is a bona fide exosomal regulator and skeletal muscle cells can release Alix-positive nano-sized extracellular vesicles, offering a new paradigm for understanding how myofibers communicate within skeletal muscle and with other organs. S-palmitoylation is a reversible lipid post-translational modification, involved in different biological processes, such as the trafficking of membrane proteins, achievement of stable protein conformations, and stabilization of protein interactions.Methods
Here, we have used an integrated biochemical-biophysical approach to determine whether S-palmitoylation contributes to the regulation of extracellular vesicle production in skeletal muscle cells.Results
We ascertained that Alix is S-palmitoylated and that this post-translational modification influences its protein-protein interaction with CD9, a member of the tetraspanin protein family. Furthermore, we showed that the structural organization of the lipid bilayer of the small (nano-sized) extracellular vesicle membrane with altered palmitoylation is qualitatively different compared to mock control vesicles.Conclusions
We propose that S-palmitoylation regulates the function of Alix in facilitating the interactions among extracellular vesicle-specific regulators and maintains the proper structural organization of exosome-like extracellular vesicle membranes.General Significance
Beyond its biological relevance, our study also provides the means for a comprehensive structural characterization of EVs. 相似文献95.
Maluquer de Motes C Clemente-Casares P Hundesa A Martín M Girones R 《Applied and environmental microbiology》2004,70(3):1448-1454
In this study, a molecular procedure for the detection of adenoviruses of animal origin was developed to evaluate the level of excretion of these viruses by swine and cattle and to design a test to facilitate the tracing of specific sources of environmental viral contamination. Two sets of oligonucleotides were designed, one to detect porcine adenoviruses and the other to detect bovine and ovine adenoviruses. The specificity of the assays was assessed in 31 fecal samples and 12 sewage samples that were collected monthly during a 1-year period. The data also provided information on the environmental prevalence of animal adenoviruses. Porcine adenoviruses were detected in 17 of 24 (70%) pools of swine samples studied, with most isolates being closely related to serotype 3. Bovine adenoviruses were present in 6 of 8 (75%) pools studied, with strains belonging to the genera Mastadenovirus and Atadenovirus and being similar to bovine adenoviruses of types 2, 4, and 7. These sets of primers produced negative results in nested PCR assays when human adenovirus controls and urban-sewage samples were tested. Likewise, the sets of primers previously designed for detection of human adenovirus also produced negative results with animal adenoviruses. These results indicate the importance of further studies to evaluate the usefulness of these tests to trace the source of fecal contamination in water and food and for environmental studies. 相似文献
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Rosina Plomp Gillian Dekkers Yoann Rombouts Remco Visser Carolien A.M. Koeleman Guinevere S.M. Kammeijer Bas C. Jansen Theo Rispens Paul J. Hensbergen Gestur Vidarsson Manfred Wuhrer 《Molecular & cellular proteomics : MCP》2015,14(5):1373-1384
Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and a fundamental component of the immune system. IgG3 represents ∼8% of the total amount of IgG in human serum and stands out from the other IgG subclasses because of its elongated hinge region and enhanced effector functions. This study reports partial O-glycosylation of the IgG3 hinge region, observed with nanoLC-ESI-IT-MS(/MS) analysis after proteolytic digestion. The repeat regions within the IgG3 hinge were found to be in part O-glycosylated at the threonine in the triple repeat motif. Non-, mono- and disialylated core 1-type O-glycans were detected in various IgG3 samples, both poly- and monoclonal. NanoLC-ESI-IT-MS/MS with electron transfer dissociation fragmentation and CE-MS/MS with CID fragmentation were used to determine the site of IgG3 O-glycosylation. The O-glycosylation site was further confirmed by the recombinant production of mutant IgG3 in which potential O-glycosylation sites had been knocked out.For IgG3 samples from six donors we found similar O-glycan structures and site occupancies, whereas for the same samples the conserved N-glycosylation of the Fc CH2 domain showed considerable interindividual variation. The occupancy of each of the three O-glycosylation sites was found to be ∼10% in six serum-derived IgG3 samples and ∼13% in two monoclonal IgG3 allotypes.Immunoglobulin G (IgG) is one of the most abundant proteins present in human serum and represents approximately three-quarters of the total serum immunoglobulin content (1). As the main mediator of humoral immunity and an important link between the adaptive and innate immune system, IgG is a fundamental component of the immune system. IgG consists of two heavy and light chains, linked by disulfide bonds. The protein can be subdivided into the antigen-binding (Fab) and the receptor-binding (Fc) region. There are four subclasses of IgG, all of which share an overall structure homology but differ slightly in their amino acid sequence; the quantity of the subclasses in human serum is as follows: IgG1 > 2 > 3 > 4 (2).IgG3 represents ∼8% of the total amount of IgG in human serum (2), and stands out from the other IgG subclasses for a number of reasons. First of all, IgG3 contains an elongated hinge region with up to a triple repeat sequence (the actual number ranging from one to three depending on the allotype (3)), which is responsible for the increased flexibility between the Fab and the Fc part, as well as the wider and more flexible angle between the two Fab arms (4, 5). This flexibility is likely the cause of the increased affinity of IgG3, compared with the other subclasses, for divalent binding to certain types of antigens (4, 6, 7). Second, IgG3 has a higher affinity for C1q, which initiates the classical complement pathway (5, 8). The interaction between IgG3 and C1q is not due to the elongated hinge region, as demonstrated by studies showing that recombinant IgG3 with an IgG1- or IgG4-like hinge sequence exhibited even greater binding affinity for C1q than wild-type IgG3 (8–10). Third, IgG3 has a higher overall affinity for the Fcγ receptors (FcγRs), through which it can influence effector cells of the innate immune system (11). The CH2 domain and hinge region of IgG3 were shown to be instrumental in binding to the high affinity FcγRI receptor (12). Finally, IgG3 generally has a shorter half-life compared with the other IgG subclasses (1 versus 3 weeks) (2). This difference was traced back to an H435R mutation that confers a positive charge at physiological pH, resulting in a decreased binding to the neonatal Fc receptor (FcRn), which is involved in recycling IgG targeted for lysosomal degradation (13). The low-efficiency FcRn-mediated transport also gives rise to decreased levels of IgG3 in mucosal tissue and impaired transport of IgG3 across the placenta (14). These properties do not hold true for all types of IgG3 since a large number of IgG3 allotypes have been described, some of which lack the H435R substitution and have a half-life and placental transport rates similar to IgG1 (13–16). IgG3 is more polymorphic than the other IgG subclasses, as evidenced by the high number of known allotypes (16). Most of the polymorphisms reside in the CH2 or CH3 domain, but the length of the hinge region can also display a high degree of variation. Depending on the number of sequence repeats, the hinge region can vary from 27 to 83 amino acid residues between different IgG3 allotypes (3, 16, 17).An N-linked complex type glycan is highly conserved and found in the CH2 domain of all IgG subclasses and allotypes. The type of glycan present at this site has been shown to influence the effector functions of IgG (18). N-glycans that lack a core fucose cause IgG to have an enhanced proinflammatory capacity through stronger binding to FcγRIIIa and FcγRIIIb (18–20). In contrast, IgG carrying sialylated N-glycans exhibits anti-inflammatory properties, likely due to increased binding affinity to C-type lectins and/or reduced binding to FcγR (18, 21, 22).O-linked glycosylation has been reported for various immunoglobulins. O-glycans are present on the hinge region of human IgA1 and IgD and mouse IgG2b (23–25). IgA1 contains nine potential sites for O-glycosylation (serine and threonine) in the hinge region, of which 3–5 are occupied, while IgD has been reported to carry between four and seven O-glycans (24–26). The O-glycosylation in the hinge of murine IgG2b was observed to protect against proteolytic digestion (23). Likewise, IgA1 was found to be more susceptible to degradation by Streptococci proteases after neuraminidase treatment (27).In this study, we report partial O-glycosylation of the human IgG3 hinge. We obtained both poly- and monoclonal IgG3 from various sources and performed proteolytic digestion with trypsin or proteinase K. NanoLC-reverse phase (RP)-ESI-ion trap (IT)-MS/MS was used to examine the resulting (glyco)peptides, revealing core 1-type O-glycans on multiple sites within the IgG3 hinge region. 相似文献
98.
Guillermo Martínez Pastur Pablo L. Peri Rosina M. Soler Stefan Schindler María Vanessa Lencinas 《Biodiversity and Conservation》2016,25(10):1843-1862
It is difficult to map and quantify biodiversity at landscape level in areas with low data availability, despite demand from decision-makers. We propose a methodology to determine potential biodiversity pattern using habitat suitability maps of the understory plant species with highest cover and occurrence frequency in the three different forests types of Tierra del Fuego (Argentina). We used a database of vascular plants from 535 surveys from which we identified 35 indicative species. We explored more than 50 potential explanatory variables to develop habitat suitability maps of the indicative species, which were combined to develop a map of the potential biodiversity. Correlation among environmental, topographic and forest landscape variables were discussed, as well as the marginality and the specialization of the indicative species. We detected differences in the niches of the species prevailing in the three forest types. The developed map of potential biodiversity uncovered hotspots of biodiversity in the ecotone of Nothofagus pumilio and N. antarctica as well as in the wettest part of the mixed N. pumilio–N. betuloides forests. It allowed thus to identify forest areas with different conservation potential and can be readily used as a decision support system for conservation and management strategies at different scales including the identification of land-use conflicts (e.g. of biodiversity with timber production and livestock) and the development of a network of protected areas, which currently does not cover the forests of highest conservation value. 相似文献
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100.
Warren W. Nichols Anthony J. Girardi Carole I. Bradt Rosina Hill Christine Cody 《Mutation research》1985,150(1-2):327-332
Human diploid fibroblasts have been transformed by ts A58 SV40. At the permissive temperature, apparent chromosome and chromatic rearrangements were observed in a high percentage of cells and the frequency of SCE increased. If the transformed phenotype returned to normal at the restrictive temperature these alterations also returned to normal levels. Chromosome banding demonstrated many apparent chromosomal rearrangements in which diffuse staining material was joining intact chromosomes end-to-end and forming pseudostructural abnormalities. Homogeneously staining regions associated with gene amplification or virus-induced alterations in the coiling and stickiness of telomeric regions are possible mechanisms. 相似文献