10.1371/journal.pone.0103743 Maarit Neuvonen Maarit Neuvonen Moutusi Manna Moutusi Manna Sini Mokkila Sini Mokkila Matti Javanainen Matti Javanainen Tomasz Rog Tomasz Rog Zheng Liu Zheng Liu Robert Bittman Robert Bittman Ilpo Vattulainen Ilpo Vattulainen Elina Ikonen Elina Ikonen Enzymatic Oxidation of Cholesterol: Properties and Functional Effects of Cholestenone in Cell Membranes Public Library of Science 2014 membrane cholesterol oxidase cholesterol content cholestenone 2014-08-26 07:22:20 Dataset https://plos.figshare.com/articles/dataset/_Enzymatic_Oxidation_of_Cholesterol_Properties_and_Functional_Effects_of_Cholestenone_in_Cell_Membranes_/1151995 <div><p>Bacterial cholesterol oxidase is commonly used as an experimental tool to reduce cellular cholesterol content. That the treatment also generates the poorly degradable metabolite 4-cholesten-3-one (cholestenone) has received less attention. Here, we investigated the membrane partitioning of cholestenone using simulations and cell biological experiments and assessed the functional effects of cholestenone in human cells. Atomistic simulations predicted that cholestenone reduces membrane order, undergoes faster flip-flop and desorbs more readily from membranes than cholesterol. In primary human fibroblasts, cholestenone was released from membranes to physiological extracellular acceptors more avidly than cholesterol, but without acceptors it remained in cells over a day. To address the functional effects of cholestenone, we studied fibroblast migration during wound healing. When cells were either cholesterol oxidase treated or part of cellular cholesterol was exchanged for cholestenone with cyclodextrin, cell migration during 22 h was markedly inhibited. Instead, when a similar fraction of cholesterol was removed using cyclodextrin, cells replenished their cholesterol content in 3 h and migrated similarly to control cells. Thus, cholesterol oxidation produces long-term functional effects in cells and these are in part due to the generated membrane active cholestenone.</p></div>