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New targeted approaches for the quantification of data‐independent acquisition mass spectrometry 下载免费PDF全文
Chih‐Chiang Tsou Alonso Barrantes‐Freer Christine Stadelmann Alexey I. Nesvizhskii Manuela Schmidt Lukas Reiter David Gomez‐Varela 《Proteomics》2017,17(9)
The use of data‐independent acquisition (DIA) approaches for the reproducible and precise quantification of complex protein samples has increased in the last years. The protein information arising from DIA analysis is stored in digital protein maps (DIA maps) that can be interrogated in a targeted way by using ad hoc or publically available peptide spectral libraries generated on the same sample species as for the generation of the DIA maps. The restricted availability of certain difficult‐to‐obtain human tissues (i.e., brain) together with the caveats of using spectral libraries generated under variable experimental conditions limits the potential of DIA. Therefore, DIA workflows would benefit from high‐quality and extended spectral libraries that could be generated without the need of using valuable samples for library production. We describe here two new targeted approaches, using either classical data‐dependent acquisition repositories (not specifically built for DIA) or ad hoc mouse spectral libraries, which enable the profiling of human brain DIA data set. The comparison of our results to both the most extended publically available human spectral library and to a state‐of‐the‐art untargeted method supports the use of these new strategies to improve future DIA profiling efforts. 相似文献
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The accessory gene regulator (agr) locus controls many of the virulence toxins involved in Staphylococcus aureus pathogenesis, and can be divided into four specificity groups. AgrC is the only group-specific receptor to mediate both intra-group
activation and inter-group inhibition. We studied the ligand-receptor recognition of the agr system in depth by using a luciferase reporter system to identify the key residues responsible for AgrC activation in two
closely related agr groups, AgrC-I, and AgrC-IV. Fusion PCR and site-directed mutagenesis were used to screen for functional residues of AgrC.
Our data suggest that for AgrC-IV activation, residue 101 is critical for activating the receptor. In contrast, the key residues
for the activation of AgrC-I are located at residues 49∼59, 107, and 116. However, three residue changes, T101A, V107S, I116S,
are sufficient to convert the AIP recognizing specificity from AgrC-IV to AgrC-I. 相似文献
65.
Camellia, comprising more than 200 species, is the type genus of the family Theaceae. Currently, the interspecies relationship of the economically important genus is still a matter of great debate and controversy. In an attempt to help settle this dispute using molecular phylogeny, we analyzed ITS sequences of 112 species of Camellia. The maximum parsimony and Bayesian trees grouped these species into eight major clades and four isolates. The current study supported the monophyly of sections Thea and Furfuracea, a merged section of Theopsis and Eriandra and the formation of section Oleifera by H, -t. Chang (Flora of Reipulicae Popularis Sinicae. Tomus 49 (3), Science Press, China). The study suggested the polyphyletic nature of the sections Camellia, Paracamellia, Pseudocamellia, and Tuberculata and the paraphyletic nature of the section Chrysantha but did not support the sectional status of the three small sections, Archecamellia, Piquetia, and Sterocarpus. We also discuss the results in terms of morphology, geographic distribution and the results from an earlier molecular phylogeny analysis. 相似文献
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Centrioles are self-reproducing organelles that form the core structure of centrosomes or microtubule-organizing centers (MTOCs). However, whether duplication and MTOC organization reflect innate activities of centrioles or activities acquired conditionally is unclear. In this paper, we show that newly formed full-length centrioles had no inherent capacity to duplicate or to organize pericentriolar material (PCM) but acquired both after mitosis through a Plk1-dependent modification that occurred in early mitosis. Modified centrioles initiated PCM recruitment in G1 and segregated equally in mitosis through association with spindle poles. Conversely, unmodified centrioles segregated randomly unless passively tethered to modified centrioles. Strikingly, duplication occurred only in centrioles that were both modified and disengaged, whereas unmodified centrioles, engaged or not, were "infertile," indicating that engagement specifically blocks modified centrioles from reduplication. These two requirements, centriole modification and disengagement, fully exclude unlimited duplication in one cell cycle. We thus uncovered a Plk1-dependent mechanism whereby duplication and segregation are coupled to maintain centriole homeostasis. 相似文献
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Wissner A Brawner Floyd MB Rabindran SK Nilakantan R Greenberger LM Shen R Wang YF Tsou HR 《Bioorganic & medicinal chemistry letters》2002,12(20):2893-2897
The syntheses and biological evaluations of 4-anilinoquinoline-3-carbonitrile analogues of the three clinical lead 4-anilinoquinazolines Iressa, Tarceva, and CI-1033 are described. The EGFR and HER-2 kinase inhibitory activities and the cell growth inhibition of the two series are compared with each other and with the clinical lead EKB-569. Similar activities are observed between these two series. 相似文献
70.
The TED (Technology, Entertainment, Design) Talks website hosts video recordings of various experts, celebrities, academics, and others who discuss their topics of expertise. Funded by advertising and members but provided free online, TED Talks have been viewed over a billion times and are a science communication phenomenon. Although the organization has been derided for its populist slant and emphasis on entertainment value, no previous research has assessed audience reactions in order to determine the degree to which presenter characteristics and platform affect the reception of a video. This article addresses this issue via a content analysis of comments left on both the TED website and the YouTube platform (on which TED Talks videos are also posted). It was found that commenters were more likely to discuss the characteristics of a presenter on YouTube, whereas commenters tended to engage with the talk content on the TED website. In addition, people tended to be more emotional when the speaker was a woman (by leaving comments that were either positive or negative). The results can inform future efforts to popularize science amongst the public, as well as to provide insights for those looking to disseminate information via Internet videos. 相似文献