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  • A detailed analysis of partial molecular volumes in DPPC/cholesterol binary bilayers.

A detailed analysis of partial molecular volumes in DPPC/cholesterol binary bilayers.

Biochimica et biophysica acta (2014-08-26)
Tsubasa Miyoshi, Max Lönnfors, J Peter Slotte, Satoru Kato
ABSTRACT

We examined the volumetric behavior of the dipalmitoylphosphatidylcholine (DPPC)/cholesterol binary bilayer system with high accuracy and more cholesterol concentrations to reveal the detailed molecular states in the liquid-disordered (Ld) phase, the liquid-ordered (Lo) phase and the gel phase. We measured the average specific volume of the binary bilayer at several temperatures by the neutral flotation method and calculated the average volume per molecule to estimate the partial molecular volumes of DPPC and cholesterol in each phase. As a result, we found that the region with intermediate cholesterol concentrations showed a more complicated behavior than expected from simple coexistence of Ld and Lo domains. We also measured fluorescence decay of trans-parinaric acid (tPA) added into the binary bilayer with more cholesterol concentrations to get further insight into the cholesterol-induced formation of the Lo phase. On the basis of these results we discuss the molecular interaction between DPPC and cholesterol molecule in the Lo phase and the manner of Ld/Lo phase coexistence.

MATERIALS
Product Number
Brand
Product Description

SAFC
Cholesterol, from sheep wool, Controlled origin, meets USP/NF testing specifications
Sigma-Aldrich
SyntheChol® NS0 Supplement, 500 ×, synthetic cholesterol, animal component-free, sterile-filtered, aqueous solution, suitable for cell culture
Sigma-Aldrich
Cholesterol, tested according to Ph. Eur.
Sigma-Aldrich
Cholesterol, Sigma Grade, ≥99%
Sigma-Aldrich
Cholesterol, powder, BioReagent, suitable for cell culture, ≥99%
Sigma-Aldrich
Cholesterol, from sheep wool, ≥92.5% (GC), powder
Supelco
Cholesterol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Deuterium oxide, 60 atom % D
Sigma-Aldrich
Deuterium oxide, 70 atom % D
Sigma-Aldrich
Deuterium oxide, filtered, 99.8 atom % D
Supelco
Cholesterol solution, certified reference material, 10 mg/mL in chloroform
Sigma-Aldrich
Deuterium oxide, 99.9 atom % D, contains 0.05 wt. % 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid, sodium salt
Sigma-Aldrich
Deuterium oxide, 99.9 atom % D, contains 0.75 wt. % 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid, sodium salt
Sigma-Aldrich
Deuterium oxide, 99.9 atom % D
Sigma-Aldrich
Deuterium oxide, 99.9 atom % D, contains 1 % (w/w) 3-(trimethylsilyl)-1-propanesulfonic acid, sodium salt (DSS)
Sigma-Aldrich
1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, ≥99% (TLC)
Sigma-Aldrich
1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, semisynthetic, ≥99%