Larry Marnett
Cellular Responses to Aldehydic Products of Lipid
Peroxidation
Aldehyde Mediators of Oxidative Damage
h y d ro p e ro x id e s , e n d o p e ro x id e s,
is o p ro s ta n e s
HO¥
O2
Ğ
+
NO
O
N
O
Fe
O–
2+
or
P LA 2
a s c o rb a te
H 2O 2
re d u c tio n
h y d ro ly sis
MDA, HNE, ONE
GSH
co n ju g a tio n
o xid a tiv e
m e ta b o lis m
P ro te in s
a p o p to s is
d e g ra d a tio n
c e ll c y c le a rre s t
p53
re p lic a tio n b lo c k
O
N
m u ta tio n
N
N
d a m a g e s ig n a lin g
N N
g e n e e x p re s s io n
Diffusible Electrophiles from Lipid Peroxidation
R
R
L ip . P e ro x.
C 5H 11
C 5H 11
O2
OOH
Ox
H
O
Ox
H
H
O
H
O
OH
C 5H 11
O
OOH
MDA
-H2O
HPNE
Red
H
H
C 5H 11
O
C 5 H 11
O
O
ONE
OH
HNE
Lipid Peroxidation-Induced DNA Damage
O
H
O
OH
N
HN
+
H 2N
O
N
N
N
N
N
N
N
M 1d G
OH
O
O
H
O
HO
H 2N
OH
N
N
N
N
HN
C 5H 11 +
N
H
N
N
N
C 5H 11
O H -H e xyl-P d G
O
NH2
H
C 5H 11
O
O
+
N
N
N
N
H 11C 5
NH
N
N
N
H -d A
N
Mutagenic Spectrum of Lipid Peroxidation-Induced Adducts
O
OH
O
HO
N
N
N
N
N
N
N
O
N
H
N
N
C 5H 11
M 1 dG
M1dGT
M1dGA
frameshifts
O H -H e xyl-P dG
OH-Hexyl-dGT
OH-Hexyl- dGA
H 11 C 5
NH
N
N
N
H -dA
N
Mutagenic Spectrum of Lipid Peroxidation-Induced Adducts
OH
O
HO
N
N
N
N
N
N
N
O
N
H
N
N
C 5 H 11
M 1d G
M1dGT
M1dGdA
frameshifts
O H -H exy l-P dG
OH-Hexyl-dGT
OH-Hexyl- dGdA
NH
N
N
N
N
dA
dA  dG
HNE and ONE Protein Adducts
O
HO
C 5H 11
H
N
L ys
O
C 5H 11
L ys
X
C y s, H is, Lys
X
R
H
R
R = C y s, H is , Ly s
C 5 H 11
O
OH
H
HNE
L ys
L ys
C 5H 11
N
–H 2O
L ys
N
C 5H 11
OH
O
C 5 H 11
H
C 5H 11
O
O
ONE
A rg
N
N
H 2N
C O 2H
plus unstable Michael adducts
N H 3+
ONE is 100x more reactive than HNE
CRT,8, 284, (1995); CRT, 16, 1598 (2003)
Enedials from Other Sources
H
C 5 H 11
O
O
ONE
O
H
P -4 50
R
H
H
O
O
O
O2
R
O
R
H
OH
P -45 0
O
O
O2
O
HO
O
O
O
HNE Induction of Apoptosis and Necrosis
CRT, 14, 1090 (2001)
Toxicity of Peroxidation-Derived Aldehydes
H
H
C 5H 11
O
OOH
HPNE
CRT, 17, 453 (2004)
H
C 5H 11
O
O
ONE
O
H
C 5H 11
OH
(R)-HNE
O
C 5H 11
OH
(S)-HNE
Depletion of GSH by HNE
Apoptosis Induced by HPNE and ONE in RKO Cells
CRT, 17, 453 (2004)
O
H
OH
4 -H N E
Cell membrane
membrane
receptor?
Mitochondria
Bcl-2
Cyt c
Apaf-1
Cas-9
Procas-3
Cas-8
Cas-3
DFF45/35
PARP
-fodrin
Protein Synthesis is Required for
HNE-Mediated Apoptosis
What transcriptional pathways are altered by HNE?
Reduced Apoptosis in p53-/- Cells Exposed to HNE
NFB Signaling
Li et al, Mol.Immunol., 41, 701 (2004)
HNE Inhibits NFB Translocation and DNA Binding
JBC, 276, 18223 (2001)
0*
Treatment
Treatment
HN E 8 0 *
HN E 6 0 *
0
HN E 4 0 *
100
HN E 2 0 *
200
0*
400
HN E
300
L uc ife ra s e ac tiv ity
(% of c on tro l)
H1299
Co ntr ol
HN E 8 0 *
HN E 6 0 *
HN E 4 0 *
HN E 2 0 *
HN E
Co ntr ol
L uc ife ra s e ac tiv ity
(% of c on tro l)
HNE Inhibits NFB-Dependent Transcription
RKO
300
250
200
RKO
150
100
50
0
HNE Modification of IB Kinase
Structure and Regulation of IB Kinase
SH
HS
SH HS
Oxidative Stress and Its Consequences
h y d ro p e ro x id e s , e n d o p e ro x id e s,
is o p ro s ta n e s
HO¥
O2
Ğ
+
NO
O
N
O
Fe
O–
2+
or
P LA 2
a s c o rb a te
H 2O 2
re d u c tio n
h y d ro ly sis
MDA, HNE, ONE
GSH
co n ju g a tio n
o xid a tiv e
m e ta b o lis m
IK
P ro te in s
a p o p to s is
d e g ra d a tio n
c e ll c y c le a rre s t
p53
re p lic a tio n b lo c k
O
N
m u ta tio n
N
N
d a m a g e s ig n a lin g
N N
g e n e e x p re s s io n
Microarray Experiments:
Dose Response, Time Course, and Chip Specifications
Doses
Times
Array Type
5, 20, and 60 µM HNE
6 and 24 h
Affymetrix
U133 Plus 2.0 Chip
~55,000 Probes
~47,000 Transcripts
CRT, 18, 1641 (2005)
Genes Up-regulated at 6 h Using Microarray
5 µM HNE
4
20 µM HNE
22
0
3
0
11
115
54520
all genes
60 µM HNE
Genes Up-regulated by HNE at 24 h
20 µM HNE
2
60 µM HNE
2
74
54597
all genes
Known Genes Upregulated by HNE
io lo g ic al R es po nse /G e ne
ntio xida nt R esp on se
em e ox yg ena se 1 ( H M O X 1 )
ut am at e-c yste in e lig as e, m od ifier sub un it (G CL M )†
A D (P)H- de pe nde nt qu in o ne o xi d ore duc ta se ( N QO 1)†
lut e c arri er fam ily 7, m em ber 1 1 (S LC7 A 11 , x CT )†
io re do xin red uc ta se 1 ( TX NR D 1)
A cc . N o .
2 0 µM H N E
6 h
6 0 µM H N E
24 h
6 0 µM H N E
N M _ 00 21 33
N M _ 00 20 61
N M _ 00 09 03
N M _ 01 43 31
N M _ 00 33 30
8 .4
3 .4
2 .5
8 .7
2 .4
9 .1
3 .1
–
6 .1
9 .3
7 .4
–
–
–
5 .9
eat Sh ock R es po nse
naJ h om ol o g, s ubf am ily A , me m ber 4 ( D na JA 4)†
N M _ 01 86 02
4 .4
3 .4
naJ h om ol o g, s ubf am ily B, m em ber 1 ( D na JB 1)†
naJ h om ol o g, s ubf am ily B, m em ber 2 ( D na JB 2)
naJ h om ol o g, s ubf am ily B, m em ber 4 ( D na JB 4)†
ea t sh oc k 7 0kD a pro te in 1A ( H SP A 1A )
ea t sh oc k 1 10 kDa pr ot e in fa mi ly (AP G-1 )
ea t sh oc k 1 05 /1 10 kDa pro te in 1 (H SP H1 )†
N M _ 00 61 45
N M _ 00 67 36
N M _ 00 70 37
N M _ 00 53 45
N M _ 01 42 78
N M _ 00 66 44
–
–
–
–
–
–
–
4 .3
4 .6
5 .7
3 .0
2 .6
9 .6
4 .4
3 .6
2 .1
3 .6
–
1 2. 1
R Stres s/A m in o Ac id/ Nut ri e nt Se nsin g Re sp on se
om oc yste in e, ER stress- in d uc ib le pro te in 1 ( H ERP )
tiva tin g tr ans crip tio n fac to r 3 (A T F3 )
lut e c arri er fam ily 3, m em ber 2 ( S LC3 A 2)
par ag in e sy nt h etase ( A S NS )
ysta th ion ase ( CT H )†
ow th arres t, D NA d ama ge- in du ci b le 34 ( G ad d3 4)†
b bl es h om olog 3 ( TR B3 )†
N M _ 01 46 85
N M _ 00 16 74
N M _ 00 23 94
N M _ 00 16 73
N M _ 00 19 02
N M _ 01 43 30
N M _ 02 11 58
–
–
–
–
–
–
–
4 .9
5 .1
2 .7
2 .8
3 .9
3 .4
3 .3
–
–
–
–
–
3 .2
–
ell Sign a lin g
suli n- in du ce d ge ne 1 ( IN S IG1 )†
AP ki na se k in ase kinas e 8 ( M A P3K 8 )
D C -lik e ki n ase 1 ( Cl k 1)
ace nt al gr ow th fa ctor ( P G F )
on ne ctiv e tissu e gro w th fac to r (CT G F )
g na l-in d uce d pro lifera tio n-ass oc ia te d 1- lik e 2 (S IP A1 L 2)†
dre no m ed ull in (A D M )
TP bi n di n g pro te in ex pres sed in s ke le tal m us cle (G EM )
be lson leu ke m ia v ira l o nc og ene hom ol o g 2 ( AB L 2)
br ob la st grow th fac to r rece pt or s ubs trate 2 (FR S 2)
A S-like w ith ou t C AAX 1 (R IT 1)
N M _ 00 55 42
N M _ 00 52 04
N M _ 00 40 71
N M _ 00 26 32
N M _ 00 19 01
N M _ 02 08 08
N M _ 00 11 24
N M _ 00 52 61
N M _ 00 51 58
N M _ 00 66 54
N M _ 00 69 12
–
–
–
–
5 .2
3 .8
–
–
–
–
–
2 .3
3 .5
3 .8
3 .5
4 .5
3 .6
3 .5
4 .9
3 .0
–
2 .6
–
2 .0
3 .2
4 .9
3 .5
–
4 .0
5 .5
–
2 .3
–
N M _ 01 56 75
N M _ 00 18 31
N M _ 00 23 92
–
–
–
5 .6
3 .2
–
3 .8
3 .9
2 .5
po ptot ic R eg ul at io n
ow th arres t, D NA d ama ge- in du ci b le 45  (G ad d4 5  )†
uster in (Cl us )†
urin e do ub le m inu te 2, hu m an ho m o lo g (HD M 2 )†
ell C ycle Re gu latio n
st rin 2 ( S ESN 2 )†
yc lin G 2 (CCN G 2)†
gu la tor of G- pro tein sign al ing 2 ( R G S)
yc lin- de pe nde nt k in ase- lik e ki n ase 3 ( C D KL 3 )
urin e-r ich bind in g e le me nt pro te in A ( P U RA )
S PY- lik e 2 ( TS PY L 2)
IO k in as e 3 (R IO K3 )
§
N M _ 03 14 59
N M _ 00 43 54
N M _ 00 29 23
N M _ 01 65 08
N M _ 00 58 59
–
3 .2
–
–
–
5 .4
2 .8
3 .7
2 .6
3 .2
–
–
3 .7
–
3 .7
N M _ 00 38 31
–
–
2 .4
ip id M eta bo lism a nd Tr a nsp ort
yt o chr ome P4 50 3 9A 1 ( CY P 39 A1 )
xy stero l b in d in g pr ot e in- lik e 1A ( O SB P L 1 A )
N M _ 01 65 93
N M _ 01 80 30
6 .1
–
7 .3
2 .3
–
–
N A P r ocess in g/ N uc le ar T r ans po rt
ol y ad en ylat ion e le me nt bi n di n g pro te in 3 ( CPEB 3 )
ow nre gu lated in o var ia n ca ncer 1 ( DO C1 )
ea th effec to r do m a in , DNA- bi n di n g do m a in 2 ( D E DD 2)
ol y ad en ylat ion e le me nt bi n di n g pro te in 4 ( CPEB 4 )†
ucle o por in li k e 1 (N U PL 1 )
u no- lik e 6, R N A bi n di n g pro tein (BR U N O L6 )
N M _ 01 49 12
N M _ 01 48 90
N M _ 13 33 28
N M _ 03 06 27
N M _ 01 40 89
N M _ 05 28 40
–
–
–
–
–
–
4 .3
3 .2
3 .0
3 .0
–
–
–
–
2 .2
2 .6
2 .1
3 .3
A cc. No .
20 µM H N E
6h
60 µM H N E
24 h
60 µM H N E
ytos ke le ta l F un ct io n/S tr uct ura l M a in te na nc e
oc oll ag en- pro lin e 4- hy dro xy las e ( P 4 H A 2)†
tin re lated pr otei n M 1 ( ARP M 1)
lde smo n 1 ( CA L D 1 )
a nsg el in (TA G L N )
ri ne pr ot e ase in hibit or H 1 ( S ERP IN H 1)
N M _ 00 41 99
N M _ 03 24 87
N M _ 00 43 42
N M _ 00 31 86
N M _ 00 12 35
–
–
2 .4
2 .3
–
3 .3
9 .2
–
–
–
–
4 .1
–
–
2 .6
ran scr ip tio na l Re gu latio n
du lt re tin a pro te in (LO C 1 53 22 2)†
rly gr ow th re sp ons e 1 (E G R 1 )
nc f in ger a nd BTB co nt ain in g pr ot ein 20 ( Z B TB 2 0)
-b ox pro tein 3 (TB X3 )
MB -b ox tra nscr ip tio n fa ct or 1 ( H BP 1 )
nc f in ger pr ot ein 10 ( ZN F 10 )
nc f in ger pr ot ein 22 7 ( ZN F 22 7)
nc f in ger pr ot ein 26 7 ( ZN F 26 7)
A M P res po nse e le m e nt bind in g pro tein 3-like 4 ( CREB P 3L 4)
AX d im eriza tio n pr ot e in 1 (M A D )
nc f in ger pr ot ein 55 4 ( ZN F 55 4)
maf o nc og en e ho m o lo g f (M A FF )†
nc f in ger pr ot ein 12 ( ZN F 12 )
ING 1 an d YY 1 bi n ding pro tein ( R Y BP )
rin ( P IR )
AD S bo x tr ans crip tio n en hanc er fac tor 2A ( M E F2A )
nc f in ger pr ot ein 27 7 ( ZN F 27 7)
nc f in ger pr ot ein 79 ( ZN F 79 )
nc f in ger pr ot ein 55 6 ( ZN F 55 6)
m o nji dom ai n c on taini n g 1C (J M JD 1C )
ix e d lin ea ge leu kem ia-tri th or ax h om ol o g 2 (M L LT 2)
x- determ ini n g r eg io n Y-b ox 18 ( S OX 1 8)
N M _ 15 36 07
N M _ 00 19 64
N M _ 01 33 60
N M _ 00 59 96
N M _ 01 22 57
N M _ 01 53 94
N M _ 18 24 90
N M _ 00 34 14
N M _ 13 08 98
N M _ 00 23 57
N M _ 15 23 03
N M _ 01 23 23
N M _ 03 32 04
N M _ 01 22 34
N M _ 00 36 62
N M _ 00 55 87
N M _ 02 19 94
N M _ 00 71 35
N M _ 02 49 67
N M _ 00 42 41
N M _ 00 59 35
N M _ 01 84 19
–
–
–
–
–
–
–
–
–
–
–
–
–
–
2 .5
–
–
–
–
–
–
–
2 .1
2 .6
2 .9
3 .1
2 .7
3 .2
2 .5
2 .2
2 .6
3 .1
2 .4
3 .7
–
–
–
–
–
–
–
2 .4
2 .9
–
–
3 .2
3 .5
–
–
–
–
–
–
–
2 .3
–
2 .2
2 .5
2 .5
2 .2
3 .1
3 .3
4 .2
–
–
2 1. 8
n H om eost asis a nd C e llu la r T ra nspo rt
a 2+ -tra nsp ortin g A TP as e, ty pe 2C , m em ber 1 ( A T P2C 1 )
ho lin erg ic r ece pt or , n ic oti n ic, a lp ha po ly pe pt ide 5 ( C H RNA 5 )
da pt or-re la ted pro te in co m p le x 4,  1 sub un it (A P4  1 )
ucle os id e d ip ho sp ha te- lin ked m o ie ty X- ty pe 9 ( N UD T 9 )
lut e c arri er fam ily 16 , me m b er 14
N M _ 01 43 82
N M _ 00 07 45
N M _ 00 65 94
N M _ 02 40 47
N M _ 15 25 27
–
–
–
–
2 .4
2 .9
2 .7
2 .2
2 .4
–
–
–
–
–
–
N A R ep ai r
bi q ui tin -c on ju ga tin g e nzy m e E 2 B (R AD 6 h omo lo g) ( U BE 2 B )
RC A- 1-i nt er actin g pro te in C -t erm in al he lic ase 1 ( BR IP 1 )
N M _ 00 33 37
N M _ 03 20 43
–
2 .3
4 .0
–
–
–
N M _ 00 35 32
N M _ 08 05 93
N M _ 00 35 18
N M _ 00 35 16
–
–
–
–
2 .7
–
–
–
–
2 .5
9 .6
2 .6
N M _ 00 19 69
N M _ 01 41 61
N M _ 00 21 61
N M _ 17 68 15
N M _ 00 05 50
–
–
–
3 .5
3 .7
–
2 .2
4 .9
–
–
2 .5
–
3 .5
–
–
iol og ical Re spo nse/G ene
ist on es
sto n e 1,
sto n e 1,
sto n e 1,
sto n e 2,
H 3c ( HI ST 1H3 E )
H 2 bk ( HI ST 1H2 BK )
H 2 bg ( HI ST 1H2 BG )
H 2a a (HI ST 2H2 AA )
r ote in Tr a nslat io n /A mi n o A c id M et ab o lism
uk ary ot ic t rans la tio n in iti atio n fac to r 5 (E IF 5 )†
ito ch on dria l ri bo som al pro te in L 1 8 ( M R PL 1 8)
o le u cine- tR NA syn th e tas e ( IAR S )
hy drof ol a te re du ct ase- lik e 1 ( DH FRL 1 )
rosin ase-r elat ed pro te in 1 (TY RP 1 )
HNE Stimulates Many Different
Transcriptional Pathways
HNE Induces Gene Expression via Many Signaling Pathways
Response
Transcription
Factors
Induced Genes
Antioxidant
Response
Nrf2
HMOX1, NQO1, GCLM, Xc–
Stress-Activated
Kinase Signaling
AP-1
stress-responsive genes
Misfolded/Damaged
Protein Response;
Amino Acid Starvation
HSFs, ATF4,
ATF6, XBP1
DnaJA4, HspA6, Hsp70B,
Gadd34, ASNS
DNA Damage
Response
p53
Mdm2, cyclin G2, p21
General Stress/
Unknown
Other/Unknown
Gadd45, Clusterin, CYP39A1
Induction of ARE Target Genes by HNE
(Real Time RT-PCR)
Induction of CYP39A1 by HNE
Increase in Protein Levels Following HNE Treatment
Activation of Stress-Responsive
Transcription Factors by HNE
Activation of NRF2
CRT, 18, 1779 (2005)
Knock-down of Nrf2 by siRNA
Nrf2 siRNA transfection
NO transfection
Negative Control siRNA
Nrf2 siRNA
-5
)
10
( x1 0
fm o l p e r u g R N A
12
8
6
percent reduction
(to neg control siRNA)
4
83%
82%
2
0
0
MG132, 5 uM
0
MG132, 5 uM
post-transfection treatment
0
MG132, 5 uM
Knock-down of Nrf2 by siRNA
-
+
-
+
-
+
none
none
control
control
Nrf2
Nrf2
MG-132 (5 mM)
siRNA
Nrf2 (68 kDa)
-tubulin (50 kDa)
nuclear protein blot
Knock-down of Nrf2 by siRNA
HO-1 mRNA (effect of Nrf2 knock-down)
NO transfection
Nrf2 siRNA
Negative Control siRNA
25
-4
)
percent expression
(to neg control siRNA)
( x1 0
fm o l p e r u g R N A
30
20
64%
15
10
46%
5
45%
0
0
HNE 15uM
HNE 45uM
0
HNE 15uM
HNE 45uM
post-transfection treatment
HNE-treatment
0
HNE 15uM
HNE 45uM
Activation of Heat-Shock Signaling by Hsp90 Modification
CRT, 19, 173 (2006)
Immunochemical Detection of HNE-Adducted Proteins
in Alcoholic Liver Disease
Microsomal
Mitochondrial
JPET, 315, 8 (2005); Chem.Res.Toxicol.18, 1324 (2005)
Protein Labeling with Azido-HNE
O
H
N3
OH
O
C H 3O
P ro te in
N3
+
P P h2
O
HN
O
H
N
O
Aaron Jacobs
O
O
NH
P rotein
H
N
O
PP h2
HN
N
H
H
N
S
3
O
O
O
NH
H
N
S
3
O
Analysis of Proteins Modified in RKO cells
0 mM
Std -FBS +FBS
50 mM 100 mM
-FBS
+FBS
Protein: 5 mg/lane
4-20% SDS-PAGE gradient gel
Nitrocellulose (0.2 mm)
Probe with S/A-HRP
Results:
Protein labeling more prominent in
cells incubated with media lacking
FBS
Most labeled proteins are high Mol.
Wt.
Andy Vila
Putative Targets of Electrophilic Lipid Peroxidation Products
HPNE, ONE, HNE
GSH
IB K in a s e
Keap1
P ro te in
P h o sp h a ta s e s
e n h a n c e d o x id a n t
s e n sitiv ity
Ca
2+
-A T P a se
s e n s itiv ity to
a p o p to s is
Low level stress  Protection
High level stress  Apoptosis
E R S tre ss
e n h a n c e d kin a s e
a c tiv a tio n
a n tio x id a n t
re s p o n s e
DNA
dam age
sig n a lin g
g e n e e x p re s s io n
m u ta tio n s
Not All Aldehydes Are Created Equal
H
O
H
C 5H 11
O
H
OH
OH
HNE
Extensive changes
in gene expression
and signaling
p53-Dependent-Apoptosis
O
OH
R
MDA
Changes
in gene expression
and signaling
p53-Dependent
G1/S and G2/M block
No Apoptosis
-Substituted-MDA
R = alkyl/aryl
No signaling changes
No Apoptosis/No Necrosis
Induction of p53-Dependent Cell Cycle Arrest and
Apoptosis by QD-MDA
H
H
O
OH
O
OH
N
N
N
Aldehyde Mediators of Oxidative Damage
h yd ro p e ro x id e s , e n d o p e ro xid e s ,
is o p ro s ta n e s
HO¥
O 2Ğ +
NO
O
N
O
F e2+ or
O–
a s co rb a te
H 2O 2
O
MDA, HNE, ONE
(C H 2 ) 3 C O 2 H
O
(D IF F U S IB L E E L E C T R O P H IL EHS )
O
H 31 C 1 5 H O
Cl
O
O
O
P
O
O
N
H 31C 15
O
O
O
O
O
O
O
O
P
O
O
H O O A -P C
C 5 H 11
O
H
O
O
N
OH
O
O
O
POPC
OH
H 31C 15
O
P
O
PGPC
O
re d u ctio n
O
N
H 31C 15
O
O
O
O
O
P
O
P E IP C
O
N
H
O
O
H 31C 15
O
O
O
O
O
P
O
N
O
Induces apoptosis in A549 cells at 100-200 nM
Uhlson et al, Chem.Res.Toxicol. 15, 896, 2002
Aldehyde Mediators of Oxidative Damage
h yd ro p e ro x id e s , e n d o p e ro xid e s ,
is o p ro s ta n e s
HO¥
O 2Ğ +
NO
O
N
O
F e2+ or
O–
a s co rb a te
H 2O 2
O
MDA, HNE, ONE
(C H 2 ) 3 C O 2 H
O
(D IF F U S IB L E E L E C T R O P H IL EHS )
O
H 31 C 1 5 H O
Cl
O
O
O
P
O
O
N
H 31C 15
O
O
O
O
O
O
O
O
P
O
O
H O O A -P C
C 5 H 11
O
H
O
O
N
OH
O
O
O
POPC
OH
H 31C 15
O
P
O
PGPC
O
re d u ctio n
O
N
H 31C 15
O
O
O
O
O
P
O
N
O
P E IP C
Protein Targets, Biological Effects?
Chuan Ji
Andy Vila
James
West
Aaron
Jacobs
CRT, 19, 173 (2006)
Acknowledgements
Carmelo Rizzo
Alan Brash
Ned Porter
Jianxin Ji
National Foundation for
Cancer Research
National Institutes of Health
HNE-Ascorbate Conjugate Found In Vivo
1.3 µM in
human plasma
Sowell et al, PNAS, 101, 17964 (2004)