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>