http:www'lipidmaps'org

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http//www.lipidmaps.org
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LIPID MAPS

• Lipid Metabolites And Pathways Strategy
• GENOMICS
• PROTEOMICS
• METABOLOMICS
• LIPIDOMICS

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Lipid Metabolites and Pathways Strategy
LIPID MAPS GOALS

• To separate and detect all of the lipids in a
  specific cell and to discover and characterize
  any novel lipids that may be present.
• To quantitate each of the lipid metabolites
  present and toquantitate the changes in their
  levels and location during cellular function.

(3) To define the biochemical pathways for each
lipid and develop lipid maps which define
the interaction networks.
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LIPID MAPS

• NIGMS Large Scale Collaborative Grant
• Glue Grant
• Mouse Macrophage cell line primary cell
• Agonist initially LPS, then oxidized LDL
• LC/Mass Spec identify new ones, quantify
• Synthesis/Characterize new ones, standards
• Bioinformatics informatics and lipid networks
• Website http//www.lipidmaps.org

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LIPID MAPS CORES
PI Edward A. Dennis - UCSD
Macrophage Biology
LC/Mass Spec
Christopher Glass - UCSD
Robert C. Murphy - Colorado
Bioinformatics
Lipid Synthesis/Characterization
Shankar Subramaniam - UCSD
Michael VanNieuwenhze - UCSD Walter Shaw - Avanti
Polar Lipids Stephen H. White - UC Irvine
Sterols
Fatty Acids/Eicosanoids
David W. Russell - UTSW
Edward A. Dennis - UCSD
Glycerophospholipids
Sphingolipids/Gangliosides
Alfred H. Merrill - Georgia Tech
H. Alex Brown - Vanderbilt

Other Lipids/Structural Lipidomics
Neutral Lipids
Christian Raetz - Duke
Robert C. Murphy - Colorado
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Phosphosphingolipids P(O2-)O-choline
Sphingomyelin
D-erythro-sphingosine
OH
O---_---
Examples of other sphingoid base backbones
NH
O
Ceramide (N-acylsphingosine)
Glycosphingolipids (examples)
D-erythro-sphinganine (dihydrosphingosine)
OH
OH
NH2
4-hydroxysphinganine (phytosphingosine)
OH
OH
HO
NH2
Gangliosides
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LC ESI MS/MS(Sullards Merrill, STKE 2000)
Cells -gt Add internal standards -gt Solvent
azeotroph (lcb, lcb-P Cer-P) -gt Extract
(SM, GlcCer, Cer, etc.) (i.e., two separate
HPLC runs/analysis)
Normal phase HPLC
Reverse phase HPLC
Mass Analyzer (Q1, Q2, Q3)
Ion Source
Inlet System
Detector
Species analyzed gtSMs, GlcCer, LacCer, Cer
Cer-P (all sphingoid base FA
variants) gtSo, Sa and 4-hydroxy-Sa (plus alkyl
chain and db variants) gtSo-P, Sa-P and other
derivatives (lysoSM, psychosine, N-methyl-)
Data System
Data out
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H
O

Palmitoyl-CoA
CH
OH
CH3(CH2)10CH2
(-)OOC
2
SCoA
Serine
()
NH
3
Serine palmitoyltransferase
3-Keto- sphinganine
O
H
CH
OH
CH3(CH2)10CH2
2
Sphingolipid biosynthesis via bioactive species
()
NH
3
NADPH
3-Ketosphinganine reductase
HO
H
Sphinganine
CH
OH
CH3(CH2)10CH2
2
()
NH
3
(Dihydro)ceramide synthase
Fatty acyl-CoA
Dihydrosphingomyelin
Sphingomyelin synthase
HO
H
CH
OH
CH3(CH2)10CH2
2
Dihydro- ceramide
Glc(Gal)ceramide synthase(s)
NH
CH3(CH2)9-19CH2
Dihydroglucosylceramide (DHGalactosylceramide)
O
NADPH
Dihydroceramide desaturase
Sphingomyelin
HO
H
CH3(CH2)10CH2
CH
OH
Ceramide
2
Sphingomyelin synthase
NH
CH3(CH2)9-19CH2
Glc(Gal)ceramide synthase(s)
O
Glucosylceramide (Galactosylceramide)
9
H
O

Palmitoyl-CoA
CH
OH
CH3(CH2)10CH2
(-)OOC
2
SCoA
Serine
()
NH
3
Serine palmitoyltransferase
3-Keto- sphinganine
O
H
CH
OH
CH3(CH2)10CH2
2
Sphingolipid turnover to bioactive
species (signals)
()
NH
3
NADPH
Sphinganine 1-phosphate
3-Ketosphinganine reductase
Sphinganine
HO
HO
H
H
CH
OPO3
CH3(CH2)10CH2
CH
OH
CH3(CH2)10CH2
2
2
Sphingosine kinase
()
()
NH
NH
3
3
(Dihydro)ceramide synthase
Fatty acyl-CoA
Dihydrosphingomyelin
Sphingomyelin synthase
HO
H
CH
OH
CH3(CH2)10CH2
2
Dihydro- ceramide
Glc(Gal)ceramide synthase(s)
NH
CH3(CH2)9-19CH2
Dihydroglucosylceramide (DHGalactosylceramide)
O
NADPH
Dihydroceramide desaturase
Sphingomyelin
HO
H
CH3(CH2)10CH2
CH
OH
Ceramide
2
Sphingomyelin synthase
NH
CH3(CH2)9-19CH2
Sphingosine 1-phosphate
Glc(Gal)ceramide synthase(s)
O
Ceramidase
Glucosylceramide (Galactosylceramide)
HO
HO
H
H
CH
OPO3
CH3(CH2)10CH2
CH
OH
CH3(CH2)10CH2
2
2
Sphingosine kinase
()
()
NH
Sphingosine
NH
3
3
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http//www.lipidmaps.org