Cell membranes contain hundreds to thousands of individual lipid species that are of structural importance but also specifically interact with proteins. Due to their highly controlled synthesis and role in signaling events sphingolipids are an intensely studied class of lipids. In order to investigate their metabolism and to study proteins interacting with sphingolipids, metabolic labeling based on photoactivatable sphingoid bases is the most straightforward approach. In order to monitor protein-lipid-crosslink products, sphingosine derivatives containing a reporter moiety, such as a radiolabel or a clickable group, are used. In normal cells, degradation of sphingoid bases via action of the checkpoint enzyme sphingosine-1-phosphate lyase occurs at position C2-C3 of the sphingoid base and channels the resulting hexadecenal into the glycerolipid biosynthesis pathway. In case the functionalized sphingosine looses the reporter moiety during its degradation, specificity towards sphingolipid labeling is maintained. In case degradation of a sphingosine derivative does not remove either the photoactivatable or reporter group from the resulting hexadecenal, specificity towards sphingolipid labeling can be achieved by blocking sphingosine-1-phosphate lyase activity and thus preventing sphingosine derivatives to be channeled into the sphingolipid-to-glycerolipid metabolic pathway. Here we report an approach using clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated nuclease Cas9 to create a sphingosine-1-phosphate lyase (SGPL1) HeLa knockout cell line to disrupt the sphingolipid-to-glycerolipid metabolic pathway. We found that the lipid and protein compositions as well as sphingolipid metabolism of SGPL1 knock-out HeLa cells only show little adaptations, which validates these cells as model systems to study transient protein-sphingolipid interactions.

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Most parts of Germany are iodine deficiency areas. Daily iodine intake may be increased by food with high iodine content. Therefore determination of iodine in different foodstuffs is of importance. Aim of our work was to develop a method for mineral waters. Besides, we wanted to find out to what extent natural mineral waters can contribute to the iodine supply of the population. The method is based on the reaction of the halogenids iodide and bromide with ethylene oxide in a sulfuric acid medium while converting into 2-iodo- and 2-bromoethanol. After extraction, the reaction products are determined by capillary gas chromatography with an electron capture detector. The method was modified for mineral waters. Single results were confirmed by ICP-MS. For mineral waters the limits of determination are 3 micrograms/L for iodide and 42 micrograms/L for bromide. The investigation of mineral waters from Hessen showed, that only few sources contain iodide in remarkable amounts. Therefore a considerable improvement of iodide intake is possible only with single mineral waters.