Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1206468 | ENA |
REPOSITORIES: ENA
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Project description:Glycosylation of proteins is one of the most common post-translational modification (PTM) and plays important regulatory functions in diverse biological processes such as protein stability or cell signaling. Accordingly, glycoproteins are also a consistent part of the human tear film proteome maintaining the proper function of the ocular surface and forming the first defense barrier of the ocular immune system. Irregularities in the glycoproteomic composition of the tear film might promote development of chronic eye diseases indicating glycoproteins as valuable source for biomarker discovery or drug target identification. The present study aimed to develop a lectin-based affinity method for the enrichment and concentration of tear glycoproteins/glycopeptides and the characterization of their specific N-glycosylation sites by high-resolution mass spectrometry (MS). For method development and evaluation, we accumulated first native glycoproteins from human tear sample pools and assessed the enrichment efficiency of different lectin column systems by 1D gel electrophoresis and specific protein stainings (Coomassie and glycoproteins). The best-performing multi-lectin column system (comprising the four lectins ConA, JAC, WGA and UEA I, termed as 4L) was applied for glycopeptide enrichment from human tear sample digests followed by MS-based detection and localization of their specific N-glycosylation sites. As main result, the present study identified in total 26 N glycosylation sites of 11 N-glycoproteins in tear sample pools of healthy individuals (n=3 biological sample pools). Amongst others, we identified tear film proteins lactotransferrin (N497 and N642, LTF), Ig heavy chain constant α-1 (N144 and 340, IGHA1), prolactin-inducible protein (N105, PIP) as well as extracellular lacritin (N105, LACRT) as highly reliable and significant N glycoproteins, which were already associated with the pathogenesis of various chronic eye diseases such as the dry eye syndrome (DES). In conclusion, the results of the present study will serve as important tear film N-glycoprotein catalogue for future studies focusing the human tear film and ocular surface-related inflammatory diseases.
2023-01-25 | PXD038280 | Pride
Project description:Marbles of Greek origin have been reported to form a brown stain layer, also known as patina or film. Early investigations discovered that this layer is formed by the two calcium oxalate minerals whewellite and weddellite. The origin of this oxalate film is, however, not clear, while one of the hypotheses is that it is a result of bacteria, fungi, or lichen growing on the surface of the marble. The present study characterizes the brown stain from one of the Parthenon marbles in Danish custody using LC-MS-based proteomics aiming to shed light on the possible biological origin of the staining layer. The marbles curated in Copenhagen are particularly interesting because the pieces have been kept indoors since before the onset of the industrial revolution, therefore they have experienced air pollution to a much lesser degree than the marbles still residing at the Acropolis.
2024-01-17 | PXD043505 | Pride
Project description:Light plays an important role in the growth and differentiation of Lentinula edodes mycelia, and mycelial morphology is influenced by light wavelengths. The blue light-induced formation of brown film on the vegetative mycelial tissues of L. edodes is an important process. However, the mechanisms of L. edodes’ brown film formation, as induced by blue light, are still unclear. Using a high-resolution liquid chromatography-tandem mass spectrometry integrated with a highly sensitive immune-affinity antibody method, phosphoproteomes of L. edodes mycelia under red- and blue-light conditions were analyzed. A total of 11,224 phosphorylation sites were identified on 2,786 proteins, of which 9,243 sites on 2,579 proteins contained quantitative information. In total, 475 sites were up-regulated and 349 sites were down-regulated in the blue vs red group. To characterize the differentially phosphorylated proteins, systematic bioinformatics analyses, including gene ontology annotations, domain annotations, subcellular localizations, and Kyoto Encyclopedia of Genes and Genomes pathway annotations, were performed. These differentially phosphorylated proteins were correlated with light signal transduction, cell wall degradation, and melanogenesis, suggesting that these processes are involved in the formation of the brown film. Our study provides new insights into the molecular mechanisms of the blue light-induced brown film formation at the post-translational modification level.
2020-10-15 | PXD016536 | Pride
Project description:The death registries of the plague epidemics of 1630, stored at the Archivio di Stato of Milano, have been interrogated via the EVA film technology (ethyl vinyl acetate film studded with crushed strong anion and cation exchangers as well as C8 resins). The EVA diskettes have been left in contact with the lower right margins of 10 different foils pertaining to the peak months of the raging disease (June through end of September) for 60-90 min and then the captured material, after elution and digestion, analyzed by mass spectrometry.
2018-03-01 | PXD008103 | Pride
Project description:A differential display experiment was done to compare genes expressed in salivary glands from unfed males, males fed in the presence of females and males fed in the absence of females. Possible differential bands were excised, cloned and sequenced. A macroarray study was done on these clones as probes. RNA from the salivary glands of ticks from the three conditions was labelled and hybridized. Two sets of primers were used in the differential display, and these same two were used in the labelling, so there are two platforms, corresponding to what primer pair was used. Two independent RNA isolations were done, Experiment 1 used one, 2 and 3 the other. Experiment 2 used exposure to X-ray film, while 1 and 3 used a phosphorimager plate. Two exposures of the X-ray film were found to give acceptable results, 1 and 5 min. These were averaged in the analysis. Keywords = salivary_gland Keywords = feeding Keywords = male Keywords = female
2010-06-09 | E-GEOD-1320 | biostudies-arrayexpress