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On the basis of HPLC it can be shown that the major cannabinoids are specifically accumulating after a shorten-ing of the light period. Chemotypes are additionally con-firmed using qPCR specific for thcaand cbdagenes. Gene transcript based chemotyping was only possible comparing the thcaand cbdatranscription level ratio at the middle of the cultivation period, while at the end differences in gene transcription ratio diminished. 1H-NMR results indicate a clear difference between both chemotypes and cultivation phases by applying pattern recognition software.Figure 1. Ratio analysis of the metabolites (Δ9-THCA/CBDA) and the relative transcription between cbdaand thca(2-(ΔΔCt), Y-ax (right) in logarithm), shown per C. sativaL. variety. A) Represent ratio analysis of chemotype I C. sativaL. var. Bedrobinol. B) Represent ratio analysis of chemo-type II C. sativaL. var. Bediol. No detectable transcription ratio could be obtained for the chemotype I C. sativaL. var. Bedrobinol in week 2 and 3 due to an absence of cbdatran-scripts. Error bars are defined by upper and lower limits for gene transcription and by SD for metabolites.L2.6RAGE receptor dimeric structure studied by hydrogen-deuterium exchange monitored by mass spectrometryEwa Sitkiewicz, Krzysztof Tarnowski, Magda Kulma, Jarosław Poznański, Michal DadlezInstitute of Biochemistry and Biophysics PAS, Warszawa, Polande-mail: Michal Dadlez <[email protected]>Introduction: Receptor for advanced glycation end prod-ucts (RAGE) is a multiligand cell surface receptor, known to bind glycoxidised form of proteins as well as many other ligands such as A‐ peptide, the S100 family of proinflam-matory cytokine-like mediators, Mac1 integrin and many others. RAGE is an important mediator of the proinflam-matory response involved in diverse pathophysiological states such as neurological disorders, stroke, amyloidosis, immune response, diabetes and inflammatory disorders, infectious disease and tumors. It is a potential therapeutic target, but in spite of significant effort its structure, ligand binding mode and signal transduction pathway has not been characterised yet on the molecular level.Methods: In the presented work we used hydrogen-deute-rium exchange and mass spectrometry to gain insight into the structural properties of exRAGE — the full extracel-lular part of the protein containing V, C1 and C2 domains and the structural consequences of its dimerisation. Results: We have identified two regions in RAGE se-quence which undergo protection when the protein dimer-ises. These regions map to the region between domains C1 and C2, indicating the molecular mechanism of stabilisa-tion of a flexible linker region. Based on this we propose a novel model of RAGE dimer and tetramer.
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Mass Spectrometry, environmental implications, Medical and Environmental Implications, Societies Joint Meeting, Polish-German Biochemical Societies