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91.
Maria Bendz Marcin Skwark Daniel Nilsson Viktor Granholm Susana Cristobal Lukas Käll Arne Elofsson 《Proteomics》2013,13(9):1467-1480
Topology analysis of membrane proteins can be obtained by enzymatic shaving in combination with MS identification of peptides. Ideally, such analysis could provide quite detailed information about the membrane spanning regions. Here, we examine the ability of some shaving enzymes to provide large‐scale analysis of membrane proteome topologies. To compare different shaving enzymes, we first analyzed the detected peptides from two over‐expressed proteins. Second, we analyzed the peptides from non‐over‐expressed Escherichia coli membrane proteins with known structure to evaluate the shaving methods. Finally, the identified peptides were used to test the accuracy of a number of topology predictors. At the end we suggest that the usage of thermolysin, an enzyme working at the natural pH of the cell for membrane shaving, is superior because: (i) we detect a similar number of peptides and proteins using thermolysin and trypsin; (ii) thermolysin shaving can be run at a natural pH and (iii) the incubation time is quite short. (iv) Fewer detected peptides from thermolysin shaving originate from the transmembrane regions. Using thermolysin shaving we can also provide a clear separation between the best and the less accurate topology predictors, indicating that using data from shaving can provide valuable information when developing new topology predictors. 相似文献
92.
Chao Ji Randy J. Arnold Kevin J. Sokoloski Richard W. Hardy Haixu Tang Predrag Radivojac 《Proteomics》2013,13(5):756-765
Searching spectral libraries in MS/MS is an important new approach to improving the quality of peptide and protein identification. The idea relies on the observation that ion intensities in an MS/MS spectrum of a given peptide are generally reproducible across experiments, and thus, matching between spectra from an experiment and the spectra of previously identified peptides stored in a spectral library can lead to better peptide identification compared to the traditional database search. However, the use of libraries is greatly limited by their coverage of peptide sequences: even for well‐studied organisms a large fraction of peptides have not been previously identified. To address this issue, we propose to expand spectral libraries by predicting the MS/MS spectra of peptides based on the spectra of peptides with similar sequences. We first demonstrate that the intensity patterns of dominant fragment ions between similar peptides tend to be similar. In accordance with this observation, we develop a neighbor‐based approach that first selects peptides that are likely to have spectra similar to the target peptide and then combines their spectra using a weighted K‐nearest neighbor method to accurately predict fragment ion intensities corresponding to the target peptide. This approach has the potential to predict spectra for every peptide in the proteome. When rigorous quality criteria are applied, we estimate that the method increases the coverage of spectral libraries available from the National Institute of Standards and Technology by 20–60%, although the values vary with peptide length and charge state. We find that the overall best search performance is achieved when spectral libraries are supplemented by the high quality predicted spectra. 相似文献
93.
Paolo Nanni Christian Panse Peter Gehrig Susanne Mueller Jonas Grossmann Ralph Schlapbach 《Proteomics》2013,13(15):2251-2255
Mass spectrometry (MS) analysis of peptides carrying post‐translational modifications is challenging due to the instability of some modifications during MS analysis. However, glycopeptides as well as acetylated, methylated and other modified peptides release specific fragment ions during CID (collision‐induced dissociation) and HCD (higher energy collisional dissociation) fragmentation. These fragment ions can be used to validate the presence of the PTM on the peptide. Here, we present PTM MarkerFinder, a software tool that takes advantage of such marker ions. PTM MarkerFinder screens the MS/MS spectra in the output of a database search (i.e., Mascot) for marker ions specific for selected PTMs. Moreover, it reports and annotates the HCD and the corresponding electron transfer dissociation (ETD) spectrum (when present), and summarizes information on the type, number, and ratios of marker ions found in the data set. In the present work, a sample containing enriched N‐acetylhexosamine (HexNAc) glycopeptides from yeast has been analyzed by liquid chromatography‐mass spectrometry on an LTQ Orbitrap Velos using both HCD and ETD fragmentation techniques. The identification result (Mascot .dat file) was submitted as input to PTM MarkerFinder and screened for HexNAc oxonium ions. The software output has been used for high‐throughput validation of the identification results. 相似文献
94.
Bertram Geinitz Lars Rehmann Jochen Büchs Lars Regestein 《Biotechnology and bioengineering》2020,117(4):999-1011
Cocultures bear great potential in the conversion of complex substrates and process intensification, as well as, in the formation of unique components only available due to inter-species interactions. Dynamic data of coculture composition is necessary for understanding and optimizing coculture systems. However, most standard online determined parameters measure the sum of all species in the reactor system. The kinetic behavior of the individual species remains unknown. Up to now, different offline methods are available to determine the culture composition, as well as the online measurement of fluorescence of genetically modified organisms. To avoid any genetic modification, a noninvasive online monitoring tool based on the scattered light spectrum was developed for microtiter plate cultivations. To demonstrate the potential, a coculture consisting of the bacterium Lactococcus lactis and the yeast Kluyveromyces marxianus was cultivated. Via partial least squares regression of scattered light spectra, the online determination of the individual biomass concentrations without further sampling and analyses is possible. The results were successfully validated by a Coulter counter-analysis, taking advantage of the different cell sizes of both organisms. The findings prove the applicability of the new method to follow in detail the dynamics of a coculture. 相似文献
95.
96.
Clarias batrachus (walking catfish) is an invasive species in Florida, renowned for its air-breathing and terrestrial locomotor capabilities. However, it is unknown how this species orients in terrestrial environments. Furthermore, while anecdotal life history information is widespread for this species in its nonnative range, little of this information exists in the literature. The goals of this study were to identify sensory modalities that C. batrachus use to orient on land, and to describe the natural history of this species in its nonnative range. Fish (n = 150) were collected from around Ruskin, FL, and housed in a greenhouse, where experiments took place. Individual catfish were placed in the center of a terrestrial arena and were exposed to nine treatments: two controls, L-alanine, quinine, allyl isothiocynate, sucrose, volatile hydrogen sulphide, pond water and aluminium foil. These fish exhibited significantly positive chemotaxis toward alanine and pond water, and negative chemotaxis away from volatile hydrogen sulphide, suggesting chemoreception – both through direct contact and through the air – is important to their terrestrial orientation. Additionally, 88 people from Florida wildlife-related Facebook groups who have personal observations of C. batrachus on land were interviewed for information regarding their terrestrial natural history. These data were combined with observations from 38 YouTube videos. C. batrachus appear to emerge most frequently during or just after heavy summer rains, particularly from stormwater drains in urban areas, where they may feed on terrestrial invertebrates. By better understanding the full life history of C. batrachus, we can improve management of this species. 相似文献
97.
Yanlan Mao Alexander L Tournier Andreas Hoppe Lennart Kester Barry J Thompson Nicolas Tapon 《The EMBO journal》2013,32(21):2790-2803
Orientation of cell divisions is a key mechanism of tissue morphogenesis. In the growing Drosophila wing imaginal disc epithelium, most of the cell divisions in the central wing pouch are oriented along the proximal–distal (P–D) axis by the Dachsous‐Fat‐Dachs planar polarity pathway. However, cells at the periphery of the wing pouch instead tend to orient their divisions perpendicular to the P–D axis despite strong Dachs polarization. Here, we show that these circumferential divisions are oriented by circumferential mechanical forces that influence cell shapes and thus orient the mitotic spindle. We propose that this circumferential pattern of force is not generated locally by polarized constriction of individual epithelial cells. Instead, these forces emerge as a global tension pattern that appears to originate from differential rates of cell proliferation within the wing pouch. Accordingly, we show that localized overgrowth is sufficient to induce neighbouring cell stretching and reorientation of cell division. Our results suggest that patterned rates of cell proliferation can influence tissue mechanics and thus determine the orientation of cell divisions and tissue shape. 相似文献
98.
Jarosław Kobak 《Biofouling》2013,29(3):141-150
Abstract The effects of several factors (shell length, exposure time, substratum orientation in space, illumination, temperature, conspecifics) upon the attachment strength (measured with a digital dynamometer) of the freshwater, gregarious bivalve Dreissena polymorpha were studied under laboratory conditions. A rapid increase in attachment strength was observed on resocart (a thermosetting polymer based on phenol-formaldehyde resin, with paper as filler) substrata during the first 4-d exposure, after which it stabilised at ca 1 N. The attachment strength increased also with mussel size. Mussel adhesion on variously oriented surfaces (vertical, upper horizontal and lower horizontal) was similar. Illumination inhibited attachment strength, as expected for a photophobic species, but only after a 2-d exposure. After 6 d, no effects of light were detected. Thus, illumination seemed to influence the attachment rate, rather than the final strength. The optimum temperature for mussel attachment was 20 – 25°C. At lower and higher temperatures (5 – 15°C and 30°C), their adhesion strength decreased. The presence of conspecifics stimulated mussel attachment strength. 相似文献
99.
In this study, the early stage of interfacial crystallisation behaviour of low molecular weight polyethylene (PE) and isotactic polypropylene (iPP) oligomer on the surface of carbon nanotubes (CNTs) with different diameters, chiralities and topography structures was studied using MD simulations. We started to simulate the effect of CNTs chirality and diameter on PE molecular chain orientation, and then the effect of CNTs topography structure on PE and iPP molecular chain orientation was investigated. Finally, some experiments were carried out to prove the simulated results. Our study shows that for CNTs with a diameter comparable with the radius of gyration (Rg) of a polymer chain, an easy orientation of PE chains along CNTs axis is observed for all the systems of the CNTs with different chiralities due to a geometric confinement effect. For CNTs with a much larger diameter, multiple orientation of PE chains is induced on its surface due to the lattice matching between graphite lattice and PE molecular chains. In this case, the chirality of CNTs dominates the orientation of graphite lattice, which determines the orientation of PE chains arrangement on CNTs surface. More importantly, it was found that the groove structure formed by CNT bundles is very useful for the stabilisation of polymer chain, and thus facilitates the orientation of molecular chain along the long axis of CNTs. As a result, a novel nanohybrid shish–kebab (NHSK) structure with CNTs acting as central shish while polymer lamellae as kebab can be successfully obtained for both PE with zigzag conformation and iPP with helical conformation. This simulation result was well supported by the experimental observation. Our study could provide not only a deep understanding of the origin of the polymer chain orientation on CNTs surface but also the guidance for the preparation of polymer/CNTs nanocomposites with novel NHSK structure. 相似文献
100.
A set of silicate ions and corresponding lithium salts have been quantum chemically (QC) simulated in a “free molecule” approach. The infrared (IR), inelastic neutron scattering (INS), and Raman spectra have been simulated and fitted to the experimentally registered ones. The complete assignment of the vibrational bands along with the intensities and potential energy distribution has been performed. The applicability of the traditionally used quasimolecule Si–O–Si model to the interpretation of bands near 440–480 cm? 1 and so-called “Boson” peak near 50 cm? 1 has been critically discussed. 相似文献