ChemBioChem
10.1002/cbic.201600245
FULL PAPER
3.36 (br s, 3H), 2.82 - 2.78 (m, 1H), 2.57 - 2.53 (m, 1H), 1.73 (h,
J = 7.5 Hz, 2H), 0.92 (t, J = 7.3 Hz, 3H). 31P-NMR (162 MHz,
D2O): -9.47 - -11.93 (m, 2P), -22.10 - - 23.33 (m, Hz). HR-ESI-
MS [M-1H]-: m/z calculated 561.0671, m/z found: 561.0681.
We gratefully acknowledge the European Research Council
(ERC Advanced Grant 339834) and the Konstanz Research
School Chemical Biology for financial support.
Keywords: epigenetics • DNA polymerase • primer extension •
5mC • modified nucleotide
6-Methyl-iso-propylamino-2-aminopurine-2´-
deoxyribonucleoside-5´-O- triphosphate (4d): Yield: 34 %
(3.4 µmol). 1H-NMR (400 MHz, d4-MeOD): 8.18 (s, 1H), 6.49 (t, J
= 6.8 Hz, 1H), 4.87 (dt, J = 3.4 Hz, 6.4 Hz, 1H), 4.42 - 4.40 (m,
2H), 4.39 - 4.26 (m, 1H), 3.49 (p, J = 1.6 Hz, 1H), 3.37 (br s, 3H),
2.93 (dt, J = 6.7 Hz, 13.5 Hz, 1H), 2.54 (ddd, J = 3.6 H, 6.3 Hz,
13.4 Hz, 1H), 1.41 (d, J = 6.8 Hz, 6H). 31P-NMR (162 MHz, d4-
MeOD): -10.44 (d, J = 22.7 Hz, 1P), -11.26 (d, J = 20.4 Hz, 1P),
-23.71 (t, J = 21.2 Hz). HR-ESI-MS [M-1H]-: m/z calculated
561.0671, m/z found: 561.0710.
[1]
a)B. H. Ramsahoye, D. Biniszkiewicz, F. Lyko, V. Clark,
A. P. Bird, R. Jaenisch, Proc. Natl. Acad. Sci. U. S. A.
2000, 97, 5237-5242; b)M. J. Ziller, F. Muller, J. Liao,
Y. Zhang, H. Gu, C. Bock, P. Boyle, C. B. Epstein, B. E.
Bernstein, T. Lengauer, A. Gnirke, A. Meissner, PLoS
genet. 2011, 7, e1002389.
[2]
[3]
Y. Xu, F. Wu, L. Tan, L. Kong, L. Xiong, J. Deng, A. J.
Barbera, L. Zheng, H. Zhang, S. Huang, J. Min, T.
Nicholson, T. Chen, G. Xu, Y. Shi, K. Zhang, Y. G. Shi,
Molecular cell 2011, 42, 451-464.
O. Ammerpohl, J. I. Martin-Subero, J. Richter, I. Vater,
R. Siebert, Biochim. Biophys. Acta 2009, 1790, 847-
862.
P. A. Jones, D. Takai, Science 2001, 293, 1068-1070.
a)F. Seela, Winter, H., Nucleosides Nucleotides 1995,
14, 129 - 142; b)M. R. Estecio, J. Gallegos, C. Vallot,
R. J. Castoro, W. Chung, S. Maegawa, Y. Oki, Y.
Kondo, J. Jelinek, L. Shen, H. Hartung, P. D. Aplan, B.
A. Czerniak, S. Liang, J. P. Issa, Genome Res. 2010,
20, 1369-1382; c)P. A. Jones, S. B. Baylin, Nat. Rev.
Genet. 2002, 3, 415-428.
6-Diethylamino-2-aminopurine-2´-deoxyribonucleoside-5´-O-
triphosphate (4e): Yield: 65 % (6.5 µmol). H-NMR (400 MHz,
1
D2O): 8.05 (s, 1H), 6.27 (dd, J = 6.2 Hz, 7.6 Hz, 1H), 4.75 (dq, J
= 2.8 Hz, 3.3 Hz, 1H), 4.28 - 4.10 (m, 3H), 3.79 - 3.69 (m, 4H),
2.76 - 2.69 (m, 1H), 2.50 (ddd, J = 3.4 Hz, 6.3 Hz, 13.9 Hz, 1H),
1.16 (t, J = 7.0 Hz, 6H). 13C-NMR (100 MHz, D2O): 158.7, 153.0,
150.8, 135.5, 112.8, 85.4, 83.0, 70.9, 65.4, 43.2, 38.7, 16.7 ppm.
31P-NMR (162 MHz, D2O): -10.02 (m, 1P), -11.40 (d, J = 19.6 Hz,
1P), -22.89 (t, J = 19.7 Hz). HR-ESI-MS [M-1H]-: m/z calculated
561.0671, m/z found: 561.0673.
[4]
[5]
6-Ethylpropylamino-2-aminopurine-2´-deoxyribonucleoside-
5´-O-triphosphate (4f): Yield: 48 % (4.8 µmol). 1H-NMR (400
MHz, d4-MeOD): 7.97 (s, 1H), 6.29 (t, J = 6.8 Hz, 1H), 4.69 (dt, J
= 3.3 Hz, 6.2 Hz, 1H), 4.24 - 4.14 (m, 2H), 4.11 - 4.08 (m, 1H),
[6]
M. M. Suzuki, A. Bird, Nat. Rev. Genet. 2008, 9, 465-
476.
3.89 (br s, 2H), 3.82 (br s, 2H), 2.73 (dt, J = 6.4 Hz, 13.6 Hz, 1H), [7]
2.34 (ddd, J = 3.5 Hz, 6.2 Hz, 13.5 Hz, 1H), 1.69 (h, J = 7.4 Hz,
A. P. de Koning, W. Gu, T. A. Castoe, M. A. Batzer, D.
D. Pollock, PLoS genet. 2011, 7, e1002384.
E. Li, T. H. Bestor, R. Jaenisch, Cell 1992, 69, 915-926.
Z. D. Smith, A. Meissner, Nat. Rev. Genet. 2013, 14,
204-220.
a)M. Rodriguez-Paredes, M. Esteller, Nat. med. 2011,
17, 330-339; b)H. Heyn, M. Esteller, Nat. Rev. Genet.
2012, 13, 679-692; c)P. W. Laird, Nat. Rev. Cancer
2003, 3, 253-266.
J. A. Law, S. E. Jacobsen, Nat. Rev. Genet. 2010, 11,
204-220.
S. H. Cross, J. A. Charlton, X. Nan, A. P. Bird, Nat.
Genet. 1994, 6, 236-244.
J. Kaput, T. W. Sneider, Nucleic Acids Res. 1979, 7,
2303-2322.
W. W. Li, L. Gong, H. Bayley, Angew. Chem., Int. Ed.
Engl. 2013, 52, 4350-4355.
G. Kubik, M. J. Schmidt, J. E. Penner, D. Summerer,
Angew. Chem., Int. Ed. Engl. 2014, 53, 6002-6006.
a)T. Wang, T. Hong, T. Tang, Q. Zhai, X. Xing, W. Mao,
X. Zheng, L. Xu, J. Wu, X. Weng, S. Wang, T. Tian, B.
2H), 1.21 (t, J = 7.0 Hz, 3H), 0.93 (t, J = 7.4 Hz, 3H). 31P-NMR
[8]
[9]
(162 MHz, d4-MeOD): -10.41 (d, J = 21.3 Hz, 1P), -11.19 (d, J =
20.5 Hz, 1P), -23.64 (t, J = 21.5 Hz). HR ESI-MS [M-1H]-: m/z
calculated 575.0827, m/z found: 575.0835.
[10]
6-Ethyl-iso-propylamino-2-aminopurine-2´-
deoxyribonucleoside-5´-O-triphosphate (4g): Yield: 27 % (2.7
µmol). 1H-NMR (400 MHz, D2O): 8.07 (s, 1H), 6.29 (t, J = 6.8 Hz,
1H), 5.27 (m, 1H), 4.75 (m, 1H), 4.24 – 4.11 (m, 3H), 3.60 (m,
2H), 2.71 (dt, J = 6.8 Hz, 13.7 Hz, 1H), 2.49 (ddd, J = 3.3 Hz, 6.4
Hz, 14.2 Hz, 1H), 1.17 (d, J = 6.8 Hz, 6H), 1.13 (t, J = 6.3 Hz,
3H). 31P-NMR (162 MHz, D2O): -10.27 (m, 1P), -11.14 (m, 1P), -
22.93 (m, 1P). HR-ESI-MS [M-1H]-: m/z calculated 575.0827,
m/z found: 575.0826.
[11]
[12]
[13]
[14]
[15]
[16]
6-Pyrrolidine-2-aminopurine-2´-deoxyribonucleoside-5´-O-
1
triphosphate (4h): Yield: 18 % (1.8 µmol). H-NMR (400 MHz,
D2O): 8.16 (s, 1H), 6.36 (t, J = 6.9 Hz, 1H), 4.86 - 4.80 (m, 1H),
4.31 - 4.27 (m, 1H), 4.25 - 4.17 (m, 2H), 4.07 - 3.55 (m, 4H),
2.80 (dt, J = 6.8 Hz, 13.8 Hz, 1H), 2.54 (ddd, J = 3.4 Hz, 6.3 Hz,
14.0 Hz, 1H), 2.11 - 2.01 (m, 4H). 31P-NMR (162 MHz, D2O): -
10.56 (d, J = 19.7 Hz, 1P), -11.26 (d, J = 19.9 Hz, 1P), -23.11 (t,
J = 18.6 Hz). HR-ESI-MS [M-1H]-: m/z calculated 559.0514, m/z
found: 559.0541.
Yuan, B. Huang, L. Zhuang, X. Zhou,
J. Am. Chem.
Soc. 2013, 135, 1240-1243; b)S. Bareyt, T. Carell,
Angew. Chem., Int. Ed. Engl. 2008, 47, 181-184.
M. Frommer, L. E. McDonald, D. S. Millar, C. M. Collis,
F. Watt, G. W. Grigg, P. L. Molloy, C. L. Paul, Proc.
Natl. Acad. Sci. U. S. A. 1992, 89, 1827-1831.
B. Akhtar-Zaidi, R. Cowper-Sal-lari, O. Corradin, A.
Saiakhova, C. F. Bartels, D. Balasubramanian, L.
Myeroff, J. Lutterbaugh, A. Jarrar, M. F. Kalady, J.
Willis, J. H. Moore, P. J. Tesar, T. Laframboise, S.
Markowitz, M. Lupien, P. C. Scacheri, Science 2012,
336, 736-739.
[17]
[18]
Acknowledgements
For internal use, please do not delete. Submitted_Manuscript