Mass spectrometry‐based profiling of phospholipids and sphingolipids in extracts from <i>Saccharomyces cerevisiae</i>

Yeast - Tập 23 Số 6 - Trang 465-477 - 2006
Xue Li Guan1, Markus R. Wenk
1Department of Biochemistry and Department of Biological Sciences, National University of Singapore, Singapore 117597

Tóm tắt

AbstractLipids are rapidly moving to centre stage in many fields of biological sciences. Lipidomics, the systems‐level scale analysis of lipids and their interacting factors, is thus an emerging field which holds great promise for drug and biomarker discovery. Here we present a mass spectrometry‐based approach for profiling of polar lipids, in particular phospholipids and sphingolipids, in Saccharomyces cerevisiae. The first step includes semi‐quantitative surveys of lipids in an untargeted fashion, which is particularly powerful for detection of changes that cannot easily be anticipated. This leads to the identification of ions with increased or decreased signal intensities. Comprehensive theoretical calculation of the masses of yeast phospholipid and sphingolipid molecular species, based on fatty acyl and headgroup heterogeneity, is next used to tentatively assign ions of interest. Subsequent targeted analysis using tandem mass spectrometry allows for characterization and quantification of phospholipids and sphingolipids. Given the high degree of conservation in pathways of lipid metabolism between different organisms, it can be expected that this method will lead to the discovery of novel enzymatic activities and modulators of known ones, particularly when used in combination with genetic and chemogenetic libraries and screens. We validated the method using the EUROSCARF library of non‐essential deletion mutants. Mutants of SCS7, a lipid hydroxylase, and SLC1, a putative acyl transferase with unknown substrate specificity, were profiled for their phospholipid and sphingolipid content. The observed changes in lipid profiles are consistent with previous observations and extend our knowledge on in vivo substrate use under permissive growth conditions. Copyright © 2006 John Wiley & Sons, Ltd.

Từ khóa


Tài liệu tham khảo

10.1016/0003-9861(72)90525-5

10.1128/jb.179.24.7611-7616.1997

Blagovic B, 2001, Lipid composition of Brewer's Yeast, Food Technol Biotechnol, 39, 175

10.1007/BF02931290

Brockerhoff H, 1963, Breakdown of phospholipids in mild alkaline hydrolysis, J Lipid Res, 35, 96, 10.1016/S0022-2275(20)40373-6

10.1146/annurev.bi.58.070189.003223

10.1146/annurev.genet.34.1.255

10.1002/(SICI)1097-0061(199812)14:16<1471::AID-YEA353>3.0.CO;2-Y

10.1128/MCB.10.5.2176

10.1002/(SICI)1097-0061(19980315)14:4<311::AID-YEA220>3.0.CO;2-B

10.1042/BJ20031949

10.1002/yea.861

10.1124/mol.65.4.813

10.1096/fj.05-5362com

10.1074/jbc.272.47.29704

10.1002/mas.20023

10.1091/mbc.E02-04-0186

Homann MJ, 1987, Coordinate regulation of phospholipid biosynthesis by serine in Saccharomyces cerevisiae, J Bacteriol, 169, 3276, 10.1128/jb.169.7.3276-3280.1987

10.1021/ac020146b

10.1034/j.1600-0854.2000.010301.x

10.1074/jbc.272.51.32566

10.1016/S0962-8924(98)01281-1

10.1016/0962-8924(96)10025-8

10.1016/0092-8674(93)90144-F

10.1016/S0021-9258(18)54011-4

Marsh D, 1990, Handbook of Lipid Bilayers.

10.1016/j.ymeth.2005.01.009

10.1073/pnas.252514899

Mullner H, 2004, Dynamics of neutral lipid storage in yeast, Acta Biochim Pol, 51, 323, 10.18388/abp.2004_3574

10.1091/mbc.10.11.3943

10.1016/S0021-9258(20)80661-9

10.1016/S0006-291X(03)01164-1

10.1016/S0021-9673(98)00021-1

10.1016/S0968-0004(00)01543-7

Ohya Y, 1986, Isolation and characterization of Ca2+‐sensitive mutants of Saccharomyces cerevisiae, J Gen Microbiol, 132, 979

10.1083/jcb.146.4.741

10.1016/S0955-0674(00)00240-4

10.1016/S0079-6603(03)75003-X

10.1038/nrd1776

10.1201/9781420027655.ch11

10.1073/pnas.0530118100

10.1128/jb.175.10.2853-2858.1993

10.1016/S0005-2736(00)00294-7

Thông tin
Thông tin xuất bản

Yeast

Tập 23 Số 6

465-477

Thông tin tác giả