Slide 1Slide 2Slide 3Slide 4Slide 5Slide 6Slide 7Slide 8Slide 9Slide 10Slide 11Slide 12Slide 13Slide 14Slide 15Slide 16Slide 17Slide 18Slide 19Slide 20Slide 21Slide 22Slide 23Slide 24Slide 25Slide 26Slide 27Slide 28Slide 29Slide 30Slide 31Slide 32Slide 33Slide 34Slide 35Slide 36Slide 37Slide 38Slide 39Slide 40Slide 41Slide 42Slide 43Slide 44Slide 45Slide 46Slide 47Slide 48Slide 49Slide 50Slide 51Slide 52Slide 53Slide 54Slide 55Slide 56Slide 57Slide 58Slide 59Slide 60Slide 61Effect of SNPs: 1R9O ~ WT with bound flurbiprophen; 1OG5 with mutations K206E, I215V, C216Y, S220P, P221A, I223L, and I224LR108flurbiprophenheme ironWT CYP 2C9 bound to flurbiprophenR108heme ironS-warfarinMutant CYP 2C9 complexed to S-warfarinCYP1A2 allele nomenclatureAllele Protein Nucleotide changes, Gene* *Position 5347 should have a T and not a C to be considered *1A.Trivial nameEffect Enzyme activity ReferencesIn vivo In vitroCYP1A2*1A CYP1A2.1 None Wild-type Normal Normal Ikeya et al, 1989 Quattrochi and Tukey, 1989 CYP1A2*1B CYP1A2.1 5347T>C Nakajima et al, 1994 Welfare et al, 1999CYP1A2*1C CYP1A2.1 -3860G>A Decreased Nakajima et al, 1999CYP1A2*1D CYP1A2.1 -2467delT Japanese patent number 05719026 Chida et al, 1999CYP1A2*1E CYP1A2.1 -739T>G Japanese patent number 05719026 Chida et al, 1999CYP1A2*1F CYP1A2.1 -163C>A Higher inducibility Japanese patent number 05719026 Sachse et al, 1999 Chida et al, 1999Han et al., 2002http://www.imm.ki.se/CYPalleles/SNPs matching: CYP1A2-03 Surrounding Sequence (GC Content=52%) CCTCAGTGTCACTGGGTAGGGGGAACTCCTGGTCCCTTGGGTATATGGAAGGTATCAGCAGAAAGCCAGCACTGGCAGGGACTCTTTGGTACAATACCCAGCATGCATGCTGTGSCAGGGGCTGACAAGGGTGCTGTCCTTGGCTTCCCCATTTTGGAGTGGTCACTTGCCTCTACTCCAGCCCCAGAAGTGGAAACTGAGATGATGTGTGGAGGAGAGASCCAGCGTTCATGTTGGGAATCTTGAGGCTCCTTTCCAGCTCTCAGATTCTGTGATGCTCAAAGGRTGAGCTCTGTGGGC(A/C)CMGGACGCAYGGTAGATGGAGCTTAGTCTTTCTGGTATCCAGCTGGGAGCCARGCACAGAACACGCATCAGTGTTTATCAAATGACTGAGGAAATGAATGARTGAATGTCTCCATCTCAACCCTCAGCCTGGTCCCTCCTKTTTTCCCTGCAGTTGGTACAGATGGCATTGTCCCAGTCTGTTCCCTTCTCGGCCACAGAGCTTYTCCTGGCCTSTGCCATCTTSTGCCTGGTATTCTGGGTGCTCAAGGGTTTGAGGCCTCGGGTCCCCAAAGGCCTGAAAAGTCCACCARAGCCATGGTo 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.dbSNP ID: rs762551 SNP500Cancer ID: CYP1A2-03 Gene: CYP1A2 SNP Region: IVS1-154C>A Note: aka CYP1A2*1F dbSNPNCBI map Ensembl map Entrez Gene1. Epoxidation of double bonds. 2. C and N hydroxylation: C-H  C-OH or N-H  N-OH3. Oxidative dealkylation: C-X-CH3  C-OH + CH2O; X= O, N, S C-NH2 C-SH4. Oxidative deamination: R-CH2-NH2  R-CH=O + NH35. N, S oxidation: R3N  R3NO ; R2SOFive reaction types of cytochrome P450OC COxidative dealkylation: C-hydroxylation followed by non-enzymatic hydrolysis of the gem-substituted adduct. H2N - - C H 2 H2N - - C H NH3 - + C H 2 = O H - O Oxidative deamination: C-hydroxylation followed by non-enzymatic hydrolysis of the gem-substituted adduct.C -X -C H3 C -X -C H2C -X -H + C H2=OX= O , N , SH -OX = O, hemiacetalX = N, gem amino hydrinX = S, thiohemiacetalOHOHOHOOOOOO123456789101112benzo[a ]pyrene1-OH6-OH3-OH*1,6-Q6,12-Q3,6-QOHHHHO9,10-DHDOOHHHOH4,5-DHDOOHHHHO7,8-DHDOOHH7,8-oxideOHH4,5-oxideOHH9,10-oxideOHHHHOOHHdiolepoxideMajor metabolites of benzo[a]pyrene by MFO system7-OH, 9-OH, 12-OH and the 4,5-Q have also been reportedOO7,8-quinoneOHHOH Arene oxide  phenol rearrangementOHHOHOHHHOHHOHStereochemistry of hydration by epoxide hydrolaseH2OHOHHHOOHHHOHHHOOHH-syn -antiHOHHHO(+)_(+)_(+)_-7,8-dihydrodiolBP diolepoxide stereo isomers(+)-anti: 7R,8S,9S,10R-BPDEABCDA>B>CSA>C>BRLowest priority atom oriented towards rear, direction ofrotation is determined on the front face. Priority is assignedby atomic number; when atoms are of equal atomic number, the atoms bonded to the atoms under consideration are considered.R, S CONVENTION+. .N H2O-H N NNNOd RHO HH N HHO HO HHHO HH N HHO HO HHNNNNd RAdducts of anti-BPDE with exocyclic amino groups of nucleobasesHeavy 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’-OH5'(+)-trans-anti--BPDE:N2-dGuo adduct at primer-template junctionOxidized angular ringDistal end of pyrene systemadduct of (-)-trans-anti-BPDE with N6-dAdo: opposite dThyd from N-Ras fragmentLooking down helix axisLooking perpendicular to helix axis5 face of dAdocis adduct of (+)-anti-BPDE at N6 of dAdo5 face of dAdo(-)-trans-anti-5-methylchrysene:dGuo adduct at the hindered bay region5'3'CH312345(+)-trans-anti-benzo[g]chrysene:dAdoInset: benzo[g]chrysene skeletonR,S-trans-anti-benzo[c]phenanthrene:dGuoS Rbenzo[c]phenanthreneclassical intercalation 5-insertion S-trans-anti-B[c]Ph 1(S) 5…C[G*]C… normal duplexclassical intercalation 3-insertion R-trans-anti-B[c]Ph 1(R) 5…C[G*]C… normal duplexBAY REGION THEORYOtransition state for opening of bay region epoxideheavy lines indicate the aromatic aromatic systemNuIf a PAH has a bay region and shows genotoxic activity, the ultimate active metabolite will be the bay region diolepoxide.OOO OCH3OOOOOOO OCH3OOOOOO OCH3Oendo (minor)exo (major)123456789aflatoxin B1AFB1AFB1-8,9-oxideCarcinogenic potency determined by:1. Intrinsic activity of ultimate active metabolite2. Proportion of metabolic pathway leading to ultimate active metabolite Aflatoxin B1 activationhuman CYP3A4OHOHHOHOO OCH3OOOHOHHOO OCH3OONHNOH2NNNReactions of AFB1-dGuo adductdepurinationhydrolysisAFB1-Fapy dGuo adductN7N9Vinyl chloride activationNONNHNONNNNNONNNNNNNNNETHENO (ADDUCTS FROM VINYL CHLORIDEN2,3-dGuo1,N2-dGuo1,N6-dAdo3,N4-dCydHNNOH2NNNH2CCHON7-ADDUCTN7-(2-oxoethyl)-dGuo+N NH3CH3CON NHOH2CH3CON NHH3CO+CH2ON NH3COHH3C+ + N2 + HO-C-hydroxylationby MFOnon-enzymaticdimethylnitrosamine (DMN)ACTIVATION OF DIMETHYLNITROSAMINEN NORR'GENERAL STRUCTUREazotic acidmonomethyl nitrosamineActivation of 2-AAF (general for amines)other amines of commercial importance2-am inonaphthalene-naphthylam ine4,4'-aminobiphenyl4,4'-diamino-3,3'-dichlorodiphenylm ethanem ethylene bis(o-chloroaniline)MOCANH2H2NNH2CH2NH2ClH2NClNNNN HON H2d RN HNN NN HON H2d