4 research outputs found

    Transferring an optimized TAP-toolbox for the isolation of protein complexes to a portfolio of rice tissues

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    Proteins are the cell's functional entities. Rather than operating independently, they interact with other proteins. Capturing in vivo protein complexes is therefore crucial to gain understanding of the function of a protein in a cellular context. Affinity purification coupled to mass spectrometry has proven to yield a wealth of information about protein complex constitutions for a broad range of organisms. For Oryza sativa, the technique has been initiated in callus and shoots, but has not been optimized ever since. We translated an optimized tandem affinity purification (TAP) approach from Arabidopsis thaliana toward Oryza sativa, and demonstrate its applicability in a variety of rice tissues. A list of non-specific and false positive interactors is presented, based on re-occurrence over more than 170 independent experiments, to filter bona fide interactors. We demonstrate the sensitivity of our approach by isolating the complexes for the rice ANAPHASE PROMOTING COMPLEX SUBUNIT 10 (APC10) and CYCLIN-DEPENDENT KINASE D (CDKD) proteins from the proliferation zone of the emerging fourth leaf. Next to APC10 and CDKD, we tested several additional baits in the different rice tissues and reproducibly retrieved at least one interactor for 81.4 % of the baits screened for in callus tissue and T1 seedlings. By transferring an optimized TAP tag combined with state-of-the-art mass spectrometry, our TAP protocol enables the discovery of interactors for low abundance proteins in rice and opens the possibility to capture complex dynamics by comparing tissues at different stages of a developing rice organ

    Deodorization of post-consumer plastic waste fractions : a comparison of different washing media

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    An important impediment to the acceptance of recyclates into a broader market is their unwanted odor after reprocessing. Different types of washing procedures are already in place, but fundamental insights into the deodorization efficiencies of different washing media are still relatively scarce. Therefore, in this study, the deodorization efficiencies of different types of plastics after washing with different media were determined via gas chromatography and mass spectrometry analysis. A total of 169 compounds subdivided into various chemical classes, such as alkanes, terpenes, and oxygenated compounds, were detected across all packaging types. Around 60 compounds were detected on plastic bottles, and around 40 were detected on trays and films. Owing to the differences in physicochemical properties of odor compounds, different deodorization efficiencies were obtained with different washing media. Water and caustic soda were significantly more efficient for poly(ethylene terephthalate) bottles with deodorization efficiencies up to 80%, whereas for polyethylene (PE) and polypropylene bottles, the washing media were relatively inefficient (around 30-40%). Adding a detergent or an organic solvent could increase deodorization efficiencies by up to 70-90% for these packaging types. A similar trend was observed for PE films having deodorization efficiencies in the range of 40-50% when washing with water or caustic soda and around 70-80% when a detergent was added.Polystyrene trays were most effectively deodorized with a detergent, achieving efficiencies up to 67%. Hence, this study shows that optimal washing processes should be tailored to specific packaging types to further improve deodorization and to eventually be able to meet ambitious European recycling targets
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