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2015 ; 112
(15
): 4558-63
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Sphingomyelin distribution in lipid rafts of artificial monolayer membranes
visualized by Raman microscopy
#MMPMID25825736
Ando J
; Kinoshita M
; Cui J
; Yamakoshi H
; Dodo K
; Fujita K
; Murata M
; Sodeoka M
Proc Natl Acad Sci U S A
2015[Apr]; 112
(15
): 4558-63
PMID25825736
show ga
Sphingomyelin (SM) and cholesterol (chol)-rich domains in cell membranes, called
lipid rafts, are thought to have important biological functions related to
membrane signaling and protein trafficking. To visualize the distribution of SM
in lipid rafts by means of Raman microscopy, we designed and synthesized an SM
analog tagged with a Raman-active diyne moiety (diyne-SM). Diyne-SM showed a
strong peak in a Raman silent region that is free of interference from intrinsic
vibrational modes of lipids and did not appear to alter the properties of
SM-containing monolayers. Therefore, we used Raman microscopy to directly
visualize the distribution of diyne-SM in raft-mimicking domains formed in
SM/dioleoylphosphatidylcholine/chol ternary monolayers. Raman images visualized a
heterogeneous distribution of diyne-SM, which showed marked variation, even
within a single ordered domain. Specifically, diyne-SM was enriched in the
central area of raft domains compared with the peripheral area. These results
seem incompatible with the generally accepted raft model, in which the raft and
nonraft phases show a clear biphasic separation. One of the possible reasons is
that gradual changes of SM concentration occur between SM-rich and -poor regions
to minimize hydrophobic mismatch. We believe that our technique of hyperspectral
Raman imaging of a single lipid monolayer opens the door to quantitative analysis
of lipid membranes by providing both chemical information and spatial
distribution with high (diffraction-limited) spatial resolution.