• Home
  • Search Results
  • Enzymatic oxidation of cholesterol: properties and functional effects of cholestenone in cell membranes.

Enzymatic oxidation of cholesterol: properties and functional effects of cholestenone in cell membranes.

PloS one (2014-08-27)
Maarit Neuvonen, Moutusi Manna, Sini Mokkila, Matti Javanainen, Tomasz Rog, Zheng Liu, Robert Bittman, Ilpo Vattulainen, Elina Ikonen
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ethyl acetate, ACS reagent, ≥99.5%
Sigma-Aldrich
Ethyl acetate, suitable for HPLC, ≥99.7%
Sigma-Aldrich
Ethyl acetate, anhydrous, 99.8%
Sigma-Aldrich
Ethyl acetate, HPLC Plus, for HPLC, GC, and residue analysis, 99.9%
Sigma-Aldrich
Ethyl acetate, puriss., meets analytical specification of Ph. Eur., BP, NF, ≥99.5% (GC)
Supelco
Ethyl acetate, analytical standard
Sigma-Aldrich
Ethyl acetate, suitable for HPLC, ≥99.8%
Sigma-Aldrich
Ethyl acetate, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
Supelco
Ethyl Acetate, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ethyl acetate, ≥99%, FCC, FG
Sigma-Aldrich
Ethyl acetate, natural, ≥99%, FCC, FG
Sigma-Aldrich
Ethyl acetate, ReagentPlus®, ≥99.8%
Sigma-Aldrich
Ethyl acetate, ACS reagent, ≥99.5%
Sigma-Aldrich
Ethyl acetate, biotech. grade, ≥99.8%
Avanti
27-hydroxy cholestenone, 27-hydroxy-4-cholesten-3-one, powder
Sigma-Aldrich
Ethyl acetate
Sigma-Aldrich
Ethyl acetate