Appendix

I. Inhibitors of Sphingolipid Biosynthesis

Jin-ichi Inokuchi

The experimental approach to deplete cellular (glyco)sphingolipids with the specific inhibitors for their biosynthetic cascade has been proved to be useful to identify functions of endogenous (glyco)sphingolipids. The site of action of each inhibitor is summarized on the biosynthesis and metabo- lism map Fig. 1. The detailed information for the inhibitors of sphingolipid synthesis and glycosphingolipid synthesis (glucosylceramide synthase) are listed in Table I and II, respectively.

>200 Structures

~ - - Ganglio series (GM3) i ~ Neolacto/Lacto series (AminoCTH)

~ - - Giobo series Isoglobo series

Sulfatide (SM4) GM4

' 7 '

ISP-1 and analogs I ~- [3 -chioro-2-alanine, ,01~.../1 L-cycloseine . ~ J ~

Serine Palmitoyl-CoA

Sulfatide (SM3)

Lactosyice ramide (LacCer)

Galactosylceramide (G alCer) Glucosylce ramide

Sphingomyelin ~ (GIcCer)

I D-PDMP and analogs I ]N-butyl-DNJ,N-buthl-DGJ ]

~Ceramide ~ Cefay~cide a lmP e

Dihydroceramide ? . . . . l~...~ / Sphingosine ~ Methylsphingosines

FumonisinB1 I ~~~---~\L.~ ' Dimethylsphingosine(DMS) ] \ , q

Sphi nganine Sphingosi ne- 1-P

& Ethanolamine-P Hexadecenal

Fig. 1. Inhibitors of (glyco)sphingolipid biosynthesis.

5 1 2 Appendix-1

Table 1. Inhibitors for sphingolipid synthesis.

Structure

..... serine palmitoyltransferase inhibitors . . . . . . . . .

Myriocin(ISP- 1 ) ,~),b) o OH il [ ooo. { /. ' ;

OH NH2

SphingofmlginB c),d~ OH NH 2

I OH

Viridiot 'unjinA d) 0 il HO COOH

,.. /- ii '-, / x .,4,. , "" .y., .... ,,COOk

o

B- chloro-L-a lanine ~)

CICH2CH(NH2)COOH

IC5o

0.3nM

3.2nM

4.7nMi

N D

ND

Cone, to be used

in cell cu l ture ,,

47nM

N D

ND

5 .... 25raM

L-cycloser ine f) N H~ .o 2 m M . . . . . . . . . . . . . . .

\\

( , . \ 0 / - N H

,,,

Ceramide synthase inhibitor Fumonis inB 1 g).h) 0.1/zM 1~50/xM

OR OH OH

i

• ., ,.,. - ~

| CH a OR CH 3 OH NH 2

R=COCH2CH(COOH)CH2COOH . . . . . . . . . . . .

a) Miyake Y, Kozulsumi Y, Nakamura S, FujilaT, Kawa~aki T. Biochem. Biophyx. Res. C):~mmun.,211,3 %(1995) b) Nakamura S, Kozu/sumi Y. Sun ~ Miyake Y, Fujita T. Kawa~ki T. J. Biol. Chem,.271,1255(1996) c) Zweerink MM, Edison AM, Wells GB. Pinto \~,, Lester RL. J. Biol. Chem. 267,2N)32( 1 ~..~)2) d) Mandala SM, Thornton RA, Frommer BR. Drei korn S, Kurtz MB. J. Antihiot., 50,339f1997) e) Gillard BK, Harrell RG, Marcus DM. Gl.vcobiology, 6,33(19%) f) Wakita H, Nishimum K,'Ibkum ~, Furukawa E Takigawa M. J. lnvesr. Dermatol.,107,336(1996) g) Wang E, Norred WR Bacon CW. Riley RT. Me,Till AH Jr. J. B#?l. Chem.. 266.14486(199.1) h) Merrill All Jr, Wang E, Gilchrist DG. Riley RI'. Adv. Lipid Res., 26,215(1 ~)3)

A p p e n d i x - I 513

Table 2. Glucosyiceramide synthase inhibitors.

Structure

P D M P analogs b D - t h r e o - P D M P ~'~ '

qa .::::==:=..

/ / . . . . . . . , \ . .... / ' - , / . / ~.

c; N ',>" ,, ~ "?, . . . . , "~ ............... ,

HN . . . . . . . . . . CH~ . .,.," \ , . . . / . . . " \ , . ,,--....\ / , . . , . \ ~ . . . . . / " . . . . . . . . . / .

0

D- t t tre o- PPt~P( P4 ) , ,,~:, ..

. J , , . . . . ,2, ,.,'=--='~,

- . . / ; ';, ¢' HN / ~ , -

"~"--"I ......... ' " " , . / " v ' " \ ....... " ...... ..--.. . . . . . . ....... . \ . . : ....... cH,

O

Ethylenedioxy-1)4 ~'~ OH O - / /

....... .,, ! ,,-:--=-==~"~ /

H~ ' , . , \ , . , " . . , , . / " ",.\. / , , " - , . / ..

i / , ",,,.../'-\, ,../',~ / ' , , - . . . . C H ~

0

D - t h r e o - P B P P ~,~ 9H

. f ........ , . . . . . -., ,i x . / - = = ~ . . ............... , .......... ..( ),

71 ~ "::' ..... S o . . . . . . . . . . .

N - a l k y l i m i n o s u g a r s N-butyl-DNJ ~,,.

U') t

N-butyl-I)GJ ~?

i i . . . . . .

=

OH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ,,,,,,

a) lnokuchi J, Radin NS. J. l.q?idRes.. 28.565(1987)

IC,

I L M

i).5

3.1

3.3

2 0

41

Conc. to be used in cell cul ture [lz M I

5"--20

5"-'20

0.01 ---0.1

5"- 160

5"--500

5"--500

b) Radin NS. Sha>'man JA, Inokuchi J. Adv. Lq,td Re.s.,26, ! 83(I 993) c) Abe A, Radin NS. Shayman JA, Wolfing LL, Zipkm RE. Sivakumar R, Ruggieri JM, Carson KG.

Ganem B. ,l. Lqmt Res.,36, 611 (! 995 ) d) Abe A., GregoIy S. Lee l... Killen PD, Brad}' RO, Kulkarni A. Shayman JA. J. ('Ira. hn,est.,105. 1563(2000) e) Jimbo M, Yamagishi K, Yamaki T. Ntmomura K, Kabayama K. Igarashi Y. Inokuchi J. ,.L Biochem.,

(Tokyo),127,485(2000) f) Platt FM, Neises GR, Dwek RA, Butters "I'D. J. Biol. ('hem.,269.8362(1994) g) Ptatt FM, Neises GR, Karlsson GB, Dwek RA. Butlers TD. ,l. Burl. ('hem..269,27108(1994)

II. The Genes in Yeast and Mammals

Yasuyuki Igarashi, Akio Kihara, and Yoshio Hirabayashi

0 H ~ S ' C ° A +- HOOC-I- CH~OH

NH~ "

palmito' ,I-CoA L-serine

II-~..]JlJI[=I lI¢. U I I ] | ( l l ~ l [ (IrC-]II-"]LI[~LM¢ ~.~[;. i I

0

NH 2 3-ketodihydrosphingosine

OH N ~ , ~ N V V V ~ O H

NHz II31~iIGt'~=1 ~ I dihydrosphingosine I~r= I ~ I'Z~ I~'-r~II

ioi~ i[¢] i ~L[i [aI~iVA ilii ~=i~l

OH

dihydroceramide

I ] l~ l~Vl , i l [ i ] [ , [ ~ 1~. ! i i l ] [ , [¢ I

OH

NH ceramide 0

t OH

N - , ~ ~ - - . ~ . ~ ~ o ~ NH O

" - - O R phosphocholine (sphingomyelin) glucose (glucosylcerarnide) galactoase (galactosylceramide) other oligosaccharides (glycosphingolipids)

m a m m a l s

OH

~ V ' ~ O H OH NH z

phytosphingosine

I IIIIIIIIIIII I~ll iI[I [ ~ v ~ ~

OH

~ O H OH NN

O phytoceramide

OH

0 m O R phosphoinositol (IPC) phosphoinositol-mannose (MIPC) phosphoinositol-mannose-phosphoinositol (M(IP)~C)

II IIII I I I

yeast

mammals, n : 1 - 8 yeast, n = 10

Fig. 1. Pathways of sphingolipid biosynthesis in yeast and mammals.

516 Appendix-H

OH 0- " ~ o - ~-o-c.~c.~.-(c.~)~

NH O

sphingomyelin 0

sphing°myelin I 1 sphingomyelinas e ............ synthase

ceramide OH OH N V ~ N V ~ ~ O H kinase O-

v V v V V v v ~ H ~ NH O

ceramide 0

ceramide synthase TI

ceramide 1-phosphate O

eramidas~ sphingosine kinase

OH OH O-

sphingosine NH 2 sphingosine 1-phosphate NH2 O

sphingosine 1°phosphate phosphohydrolase [

O O + r"o-~,-o-

NH~ ~ hexadecenal phosphoethanolamine

Fig. 2. Sphingomyelin metabolic pathways.

Appendix-H 517

Table 1. Genes involved in de novo sphingolipid synthesis and sphingomyelin metabolism.

Gene S. cerev i s iae H o m o sap iens

Serine palmitolytransfevase

3 - K e t o d i h y d r o s p h i n g o s i n e reductase

LCB1/1SC2/END8/YMR296c (M63674) SPIZC1/LCB1 (Y08685) LCB2/TSC1/SCS1/YDR()62w (I B3931 ) SPILC2/LCB2 (Y08686) TSC3/YBRO85c-a (Z35955)

TSClO/YBR265w (Z36134) FVTI (X63657)

Ceramide synthase LAG1/YHLO03c (AY558514) LAC1/YKLO08c (Z28008) LIP 1 / YMR298w ( X 80836)

Sphingolipid C-4 hydroxylase/ SUR2/SYR2/YDR297w (l.r07171 ) b A4-desaturase

Sphingomyelin synthase/ IPC synthase

AURI/YKLOO4w (U49090)

Sphingomyelinase,qnositol phosphosphingolipid phospholipase C

IS(.'I/YEROI9w ([i 1.8778)

Ceramidase YPCt/YBRI83w (AF 191745) YDCI/YPLO87w (AF214455)

Sphingosine kinase

Sphingosine 1 phosphate phoshohydrolase

LCB4/Y()R t 71c (Z75078) LCB5/IZR260w (U 17244)

LCB3/ES'R2/LBP 1/Ell. 134w (Z'194 ! O) E~R3/LBP2/Yh'R053c (Z28278)

LASS1/UOG1/LAG 1 (AFI05005) LASS2/TRH3 (BC010032) LASS3 (NM_178842) a LASS4/I'RH1 (NM_024552) LASS5/TRH4 (BC032565) LASS6 (NM_203463)

DES1 (AF002668) c DES2 (NM_206918) d

SMS1 (AB 154421 ) SMS2 (NM_ 152621 )

SMI-'I) 1/ASM (M59916) SMPD2/nSMase (AJ222801) SMPD3/nSMase 2 (AJ 2504(.~3) alk-S,Vlase (AY230663)

ASAItl/ASAIt/AC (U70063) ASA H2/NCDase/Mito- CDase

(AF449759, AF250847) e ASAH3/ACERI (AF347024) PHCA/APHC (AF214454)

SPHK1 (BC030553, NM_021972, AF200328) e

SPHK2 (BC006161, AF245447)e

SPPI/SGPPI (AJ293294) St'P2/SGPP2 (AF542512)

~ "~ 8'*5 Sphingosine 1 -phosphale DPL1/BST1/YDR294c (U51031 ) SI L/SG[ L (AI1 ,~8 .... ) lyase

f'eramide kinase not kmnd CERK (AB079066)

a There is no report describing the enzyme activity at present. h Sphinsoshine C-4 hydroxylase, which prefers sphinganine to dihydroceramide. ': Dihydroceramide A4-desaturase. d Dihydroceramide C-4 hydroxylase/A4-desaturase, a bifunctional enzyme. e Variants that differ in their N-terminal lengths exist.

Keyword Index

A

acid sphingomyelinase 233 acute lung injury (ALl) 403 adhesion 253 aggregation 329 Alzheimer' s disease 309, 319 amyloid beta protein 309, 319 angiogenesis 183 apoptosis 69, 167, 285, 337,

453 ATP binding cassette transporter

263 axon 253 axonal degeneration 329

B

B cell receptor 245 B cell survival 245 bacterial prototype 483

C

C4-hydroxylase 57 calcium 285 cancer 363 carcinogenesis 363 caveolae 233 caveolin 1,2 233 CD38 245

cell migration 415 ceramide 49, 57, 69, 153, 167,

183,337 ceramide glucosyltransferase

263,443,453 ceramide kinase 207,337 ceramide synthase 153,345 ceramide 1-phosphate 207 CERKL 337 CERT 107 conditional knockout 443 crystal structure 483 cytoskeleton 403

D

delta4-desaturase 57 detergent-insoluble microdomain

233,273 detergent-resistant mincrodomain

(DRM) 233,273 dihydroceramide synthase 49 disease 3, 25 Drosophila melanogaster 453

E

Edg receptor 385,415 endothelium 403

530 Keyword Index

FAN (factor associated with N-SMase activation)

FFAT 107 flip-flop 95 folate 345 fumonisin 345

167

G

galactosylceramide ganglioside 309 ganglisoide GM3 Gaucher disease gene-midified mice glucosylceramide

83

273 83

443 83,453

glucosylceramide synthase 263,443,453

glucosyltransferase 453

glycosphingolipidoses glycosphingolipids

263, 443,

285 295,319

G-protein coupled receptors (GPCRs) 385, 415

guidance 253

H

hereditary sensory neuropathy (HSN) 329

L

LASS gene 49 lateral segregation 123 lipid phosphate phosphatase

427 lipid probe 475 lipid raft 233,253,273,475 long-chain base 95, 219 lysenin 475

M

membrane domain(s) 245,475 microdomain 309 Mss4 463 multidrug resistance 263 myriocin 463

N

neural tube defects 345 neuron 253 neutral ceramidase 183 neutral sphingomyelinase 233 Niemann Pick disease (NPD)

167

O

organelle 123

imino sugars 83 immunity 385 inflammation 285 insulin receptor 273 insulin resistance 273 intestinal epithelial cells ISP-1 463

57

P

pathway visualization PDMP 83 permeability 403 phospholipids 285 photoreceptors 337 phytoceramide 57 phytosphingosine 95

493

Keyword Index 531

Pkh 1 141,463 Pkh2 141 plasma membrane 233 platelets 427 pleckstrin homology (PH) domain

107,207 pore-forming toxin 475 protein transport 319, 329 purification 49 pyridoxal 5'-phosphate 219

sphingolipid structure 3 sphingolipid synthesis 153 sphingolipid-containing bacteria

483 sphingolipidomic 493 sphingomyelin 69, 107 sphingomyelin synthase 69 sphingomyelinase 167 sphingosine 95, 141, 197 sphingosine kinase 197

R

rab GTPases 295 Rac 415 raft 233,253,273,475 reaction mechanism 483 regulation 25 Retinitis Pigmentosa 337

S

S 1P lyase (SPL) 219 S 1P receptor 183 Saccharomyces cerevisiae secretase 319 seed 309 serine palmitoyltransferase

sphingosine- 1-phosphate (S 1P) 183, 197, 219, 385, 403, 415,427

SphingoViz 493 SPTLC1 329 START 107 stress 25

141

tissue-specific distribution trafficking 295 translocation 123 transport and sorting 123

V

vascular biology 427 25,153,443,463,483

Sgpll 219 Slil 463 sphingolipid(s) 123,263,385 sphingolipid bases 153 sphingolipid biological functions

3 sphingolipid biosynthesis 25 sphingolipid metabolism 3 sphingolipid metabolites 363 sphingolipid signaling 463 sphingolipid storage diseases

295

vascular endothelial cells vascular smooth muscle cells

427

Y

Ypkl 463

zebrafish early development 183

57

427