Effect of SNPs: 1R9O ~ WT with bound flurbiprophen; 1OG5 with mutations K206E, I215V, C216Y, S220P, P221A, I223L, and I224L

R108 flurbiprophen heme iron WT CYP 2C9 bound to flurbiprophen

R108 heme iron S-warfarin Mutant CYP 2C9 complexed to S-warfarin

CYP1A2 allele nomenclature AlleleProteinNucleotide changes, Gene * Gene *Position 5347 should have a T and not a C to be considered *1A. Trivial name EffectEnzyme activityReferences In vivoIn vitro CYP1A2*1ACYP1A2.1NoneWild- type Normal Ikeya et al, 1989 Quattrochi and Tukey, 1989 CYP1A2*1BCYP1A T>C Nakajima et al, 1994 Welfare et al, 1999 CYP1A2*1CCYP1A G>A Decreased Nakajima et al, 1999 CYP1A2*1DCYP1A delT Japanese patent number Chida et al, 1999 Chida et al, 1999 CYP1A2*1ECYP1A T>G Japanese patent number Chida et al, 1999 Chida et al, 1999 CYP1A2*1FCYP1A C>A Higher inducibility Japanese patent number Sachse et al, 1999 Chida et al, 1999 Han et al., 2002 Sachse et al, 1999 Chida et al, 1999 Han et al.,

SNPs matching: CYP1A2-03 Surrounding Sequence (GC Content=52%) CCTCAGTGTCACTGGGTAGGGGGAACTCCTGGTCCCTTGGGTAT ATGGAAGG TATCAGCAGAAAGCCAGCACTGGCAGGGACTCTTTGGTACAATA CCCAGCAT GCATGCTGTGSCAGGGGCTGACAAGGGTGCTGTCCTTGGCTTCC CCATTTTG GAGTGGTCACTTGCCTCTACTCCAGCCCCAGAAGTGGAAACTGA GATGATGT GTGGAGGAGAGASCCAGCGTTCATGTTGGGAATCTTGAGGCTCC TTTCCAGC TCTCAGATTCTGTGATGCTCAAAGGRTGAGCTCTGTGGGC(A/C )CMGGACG CAYGGTAGATGGAGCTTAGTCTTTCTGGTATCCAGCTGGGAGCC ARGCACAG AACACGCATCAGTGTTTATCAAATGACTGAGGAAATGAATGART GAATGTCT CCATCTCAACCCTCAGCCTGGTCCCTCCTKTTTTCCCTGCAGTT GGTACAGA TGGCATTGTCCCAGTCTGTTCCCTTCTCGGCCACAGAGCTTYTC CTGGCCTS TGCCATCTTSTGCCTGGTATTCTGGGTGCTCAAGGGTTTGAGGC CTCGGGTC CCCAAAGGCCTGAAAAGTCCACCARAGCCATGGS RA/CMYR KYS R To link to the SNP in the Genewindow genome browser, click on the red SNP. To view one of the other SNPs in this sequence, click on its IUPAC code. Some SNPs in this sequence are not currently in the database.IUPAC code dbSNP ID: rs SNP500Cancer ID: CYP1A2- 03 Gene: CYP1A2 SNP Region: IVS1-154C>A Note: aka CYP1A2*1F CYP1A2IVS1-154C>A dbSNP NCBI map Ensembl mapdbSNP NCBI map Ensembl map Entrez GeneEntrez Gene

1. Epoxidation of double bonds. 2. C and N hydroxylation: C-H  C-OH or N-H  N-OH 3. Oxidative dealkylation: C-X-CH 3  C-X-CH 2 -OH  CXH + CH 2 O; X= O, N, S C-X-CH 2 -NH 2 C-X-CH 2 -SH 4. Oxidative deamination: R-CH 2 -NH 2  R-CH(OH)NH 2  R-CH=O + NH 3 5. N, S oxidation: R 3 N  R 3 N  O ; R 2 S  O Five reaction types of cytochrome P450

Oxidative dealkylation: C-hydroxylation followed by non-enzymatic hydrolysis of the gem-substituted adduct. Oxidative deamination: C-hydroxylation followed by non-enzymatic hydrolysis of the gem-substituted adduct. C-X-CH 3 C-X-CH 2 C-X-H + CH 2 =O X=O, N, S H-O X = O, hemiacetal X = N, gem amino hydrin X = S, thiohemiacetal

Arene oxide  phenol rearrangement

Stereochemistry of hydration by epoxide hydrolase H2OH2O

BP diolepoxide stereo isomers (+)-anti: 7R,8S,9S,10R-BPDE

Adducts of anti-BPDE with exocyclic amino groups of nucleobases Heavy lines show the aromatic π-system conjugated with, and stabilizing the incipient positive charge resulting from attack at epoxide ring. This situation favors addition adjacent to aromatic ring.

3’-OH

Oxidized angular ring Distal end of pyrene system

adduct of (-)-trans-anti-BPDE with N 6 -dAdo: opposite dThyd from N-Ras fragment Looking down helix axis Looking perpendicular to helix axis 5 face of dAdo

cis adduct of (+)-anti-BPDE at N 6 of dAdo 5 face of dAdo

(–)-trans-anti-5-methylchrysene:dGuo adduct at the hindered bay region 5' 3'

(+)-trans-anti-benzo[g]chrysene:dAdo Inset: benzo[g]chrysene skeleton

R,S-trans-anti-benzo[c]phenanthrene:dGuo SR

classical intercalation 5-insertion S-trans-anti-B[c]Ph 1(S) 5…C[G*]C… normal duplex classical intercalation 3-insertion R-trans-anti-B[c]Ph 1(R) 5…C[G*]C… normal duplex

BAY REGION THEORY O  transition state for opening of bay region epoxide heavy lines indicate the aromatic  aromatic system  Nu If a PAH has a bay region and shows genotoxic activity, the ultimate active metabolite will be the bay region diolepoxide.

Aflatoxin B 1 activation human CYP3A4

Reactions of AFB 1 -dGuo adduct depurination hydrolysis

AFB 1 -Fapy dGuo adduct N7 N9

AFB 1 -Fapy dGuo adduct DNA helix in ribbon form DNA helix omitted

Vinyl chloride activation

+ NO N N HN O N N N N N O N N N N N N N N N ETHENO (  ADDUCTS FROM VINYL CHLORIDE N 2,3-  dGuo 1,N 2 -  dGuo 1,N 6 -  dAdo 3, N 4 -  dCyd HN N O H 2 N N N H 2 C CH O N7-ADDUCT N7-(2-oxoethyl)-dGuo

ACTIVATION OF DIMETHYLNITROSAMINE GENERAL STRUCTURE azotic acid monomethyl nitrosamine

Activation of 2-AAF (general for amines) other amines of commercial importance

Nitrenium ion adducts C8 adduct of AAF C1 adduct of AAF C8 adduct 1-aminopyrene

NMR structure of 1-aminopyrene adduct at C8 of dG opposite dA modified dG pyrene

Activation of nitroaromatics-multiple pathways, MFO and nitroreductase Other important environmental nitro- PAH

Deamination via diazotization reaction

Deamination by bisulfite reaction

Direct acting mutagens

P450 catalytic cycle “compound I” “compound 0”

Proposed mode of action of PCBs

2O 2 + 2H + H 2 O H M n+ M (n+1)+ + HO - + HO SOD - Hydroxyl radical from action of superoxide dismutase followed by Fenton chemistry Fenton reaction

Reductive dehalogenation of carbon tetrachloride

LIPID PEROXIDATION RADICAL CHAIN

α β β-CLEAVAGE TO UNSATURATED ALDEHYDES

Formation of Malondialdehyde

OH O malondialdehyde (enol form) N N N O N N dR 1,N 2 -propeno dG O N OH N N O N N dR NHO N N O N N dR acrolein CH 3 O NH 3 C OH N N O N N dR crotonaldehyde O OH N OH N N O N N dR HO 4-hydroxy-2-nonenal 4 isomers O OH O 4-hydroxy-2,3-epoxynonanal N O N N dR N N N O N N dR N N O 1,N 2 -etheno dG 4 isomers + + 1,N 2 -propano dG, two isomers OH (M 1 G)

 dGuo from initial 2,3-epoxy-4-hydroxynonanal adducts

HN N N N O dR H 2 N HN N N H N O dR H 2 N O N N NH O dR H 2 N N O H 2 N H 2 N O NH dR 2,2-diamino-4-[(2’-deoxyribosyl)- amino]-5(2H)-oxazolone dZ H 2 O 1-electron oxidation products of dGuo NH N N H N dR H 2 N O O -e - -e -

HN N N N H 2 N O dR HN N N H N H 2 N O dR O HN N NH NH H 2 N O dR H O N N NH dR O H 2 N O N NH dR O H 2 N H 2 N HO HO HO 8-oxodGuo FapyGuo Hydroxyl radical oxidation of dGuo 2,2-diamino-4-[(2’-deoxyribosyl)- amino]-5(2H)-oxazolone dZ

N N NH 2 O dR OH OH H H HN N O O dR OH OH CH 3 H N N NH 2 OH O dR 5-hydroxy dCyd 5,6-dihydroxy-5,6-dihydro dCyd dThyd glycol Products of Hydroxyl Radical Oxidation of Bases

Oxidation of 8-oxo-Gua by peroxynitrite (ONOO - )

Formation of base propenal Reaction of base propenal with dGuo to form M 1 G

M 1 G FROM BASE PROPENAL

Cation radicals from 1-electron oxidation of BP and 6-MePB benzo[a]pyrene cation radical6-methylbenzo[a]pyrene cation radical

Adducts of the cation radicals of BP and 7,12-DMBA with dAdo and dGuo

Pathway for formation of adducts of dGuo from the cation radical of DMBA

Pathway to adducts of dAdo from 1-electron oxidation of dibenzo[a,l]pyrene

“ene-diyne” class of antineoplastic drugs H1´ abstraction from deoxyribose From: Chem. Rev. 1998, 98,