Dataset Information

Phase Composition Control in Microsphere-Supported Biomembrane Systems.

ABSTRACT: The popularization of studies in membrane protein lipid phase coexistence has prompted the development of new techniques to construct and study biomimetic systems with cholesterol-rich lipid microdomains. Here, microsphere-supported biomembranes with integrated ?-helical peptides, referred to as proteolipobeads (PLBs), were used to model peptide/protein partitioning within DOPC/DPPC/cholesterol phase-separated membranes. Due to the appearance of compositional heterogeneity and impurities in the formation of model PLB assemblies, fluorescence-activated cell sorting (FACS) was used to characterize and sort PLB populations on the basis of disordered phase (Ld) content. In addition, spectral imaging was used to assess the partitioning of FITC-labeled ?-helical peptide between fluorescently labeled Ld phase and unlabeled ordered phase (Lo) phase lipid microdomains. The apparent peptide partition coefficient, Kp,app, was measured to be 0.89 ± 0.06, indicating a slight preference of the peptide for the Lo phase. A biomimetic motif of the Lo phase concentration enhancement of the biotinyl-peptide ligand display in proteolipobeads was also observed. Finally, peptide mobility was measured by FRAP separately in each lipid phase, yielding diffusivities of 0.036 ± 0.005 and 0.014 ± 0.003 ?m2/s in the Ld and Lo phases, respectively.

SUBMITTER: Fried ES

PROVIDER: S-EPMC5568755 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature

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Phase Composition Control in Microsphere-Supported Biomembrane Systems.

Fried Eric S ES Li Yue-Ming YM Gilchrist M Lane ML

Langmuir : the ACS journal of surfaces and colloids 20170314 12

The popularization of studies in membrane protein lipid phase coexistence has prompted the development of new techniques to construct and study biomimetic systems with cholesterol-rich lipid microdomains. Here, microsphere-supported biomembranes with integrated α-helical peptides, referred to as proteolipobeads (PLBs), were used to model peptide/protein partitioning within DOPC/DPPC/cholesterol phase-separated membranes. Due to the appearance of compositional heterogeneity and impurities in the ...[more]

PMID: 28198634

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Dataset Information

Phase Composition Control in Microsphere-Supported Biomembrane Systems.

Publications

Phase Composition Control in Microsphere-Supported Biomembrane Systems.

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