Biology Reference
In-Depth Information
Abstract
Lipid droplets (LD) are in the spotlight of lipid research because of the link of lipid
storage to health and disease and the just incipient understanding of their involve-
ment in cellular processes apart from nonpolar lipid metabolism. Yeast is an excel-
lent model organism to study the lipidome and proteome of LD under different
environmental conditions and to address new aspects of LD biology and chemistry.
In this chapter, we describe a versatile protocol for the isolation of LD at high purity
and address specific demands for handling different yeast species. Moreover, we dis-
cuss the analysis of the LD proteome and lipidome based on standard methods
such as thin layer chromatography (TLC), gas liquid chromatography (GLC), mass
spectrometry (MS) as well as GLC/MS. Finally, we point out similarities and dispar-
ities of LD proteome and lipidome from the three different yeasts
Saccharomyces
cerevisiae
,
Yarrowia lipolytica
, and
Pichia pastoris
.
Abbreviations
ER
endoplasmic reticulum
FA
fatty acids
GLC
gas liquid chromatography
HPLC
high performance liquid chromatography
LD
lipid droplets
MS
mass spectrometry
nLC
nanoliquid chromatography
PC
phosphatidylcholine
PE
phosphatidylethanolamine
PI
phosphatidylinositol
PS
phosphatidylserine
RT
room temperature
SE
steryl esters
TG
triacylglycerols
TLC
thin layer chromatography
INTRODUCTION
Yeast is a well-established model organism to study the synthesis and turnover of
nonpolar lipids, which are inevitably linked to a very specific cellular compartment
named lipid droplets (LD) (
Athenstaedt & Daum, 2006; Zweytick, Athenstaedt, &
Daum, 2000
). They are small spherical organelles with an approximate diameter
of 400 nm in
Saccharomyces cerevisiae
. Increasing interest in LD biochemistry
and biology is not only due to their universal occurrence in almost all kingdoms
of life, but also to the involvement of lipid storage in health and disease. Moreover,
our understanding of LD participation in cellular processes apart from nonpolar lipid
turnover is steadily increasing but nevertheless just at its infancy.
