A versatile approach towards regioselective platinated DNA sequences.
Undesired N(7) platination of 2'-deoxyguanosine residues at predetermined sites in an oligodeoxynucleotide (ODN) sequence is prevented by applying the sterically demanding diphenylcarbamoyl (DPC) as an O(6)-protecting group. The presence of a base-labile oxalyl linker between the immobilized 3'-nucleotide and controlled pore glass (CPG) allows cleavage of the protected ODN from the support and leaves DPC protection unaffected. This method provides an ODN with specifically blocked guanine-N(7) sites for platination. In the hexanucleotides prepared in this study, 5'-GGBGGT-3'(for B=T, C and A), a platinum GG adduct is introduced at G4,G5. These site-specific platinated hexamers were isolated in a yield of 65 %, and were fully characterized by using reversed-phase HPLC (high performance liquid chromotography), LCMS (liquid chromatography-mass spectrometry), MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry), PAGE (polyacrylamide gel electrophoresis) and Maxam-Gilbert sequencing analysis.
Heetebrij
Robert J
RJ
Leiden Institute of Chemistry Gorlaeus Laboratories, Leiden University, P.O. Box 9502, The Netherlands.
de Kort
Martin
M
Meeuwenoord
Nico J
NJ
den Dulk
Hans
H
van der Marel
Gijs A
GA
van Boom
Jacques H
JH
Reedijk
Jan
J
D001483:Base Sequence; D002219:Carbamates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005898:Glass; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D009944:Organoplatinum Compounds / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013237:Stereoisomerism
D002219:Carbamates; D003432:Cross-Linking Reagents; D018736:DNA Adducts; D009838:Oligodeoxyribonucleotides; D009944:Organoplatinum Compounds; D004247:DNA; C070766:carbamic acid