Detection of the sites of alkylation in DNA and polynucleotides by laser Raman spectroscopy.
A laser Raman study of the alkylation of calf thymus DNA, poly(dG)-poly(dC) and poly(dA)-(dT) has been made using two water soluble alkylating agents: an antitumor drug, the difunctional methyl nitrogen mustard (HN2), which froms interstrand cross-links, and the dimethyl nitrogen half mustard (HN1). When an excess of the alkylating agent was used, the observed Raman frequencies due to the guanine ring modes in DNA and poly(dG)-poly(dC) changed virtually quantitatively to those of 7-methylguanosine (7-Me-Guo) showing that essentially all of the guanine bases were alkylated in the N-7 position. Furthermore, this alkylated DNA formed a stable double helical complex at neutral pH in which the alkylated guanine residues are in the keto form. No changes in the Raman bands of any of the other bases were observed in alkylated DNA. The DNA double helix, completely alkylated in at the N-7 position of guanine, melts about 35 degrees C below that of the native DNA. Upon melting, the alkylated guanine changes from the keto to the zwitterionic form.
D000478:Alkylation; D001665:Binding Sites; D055598:Chemical Phenomena; D002621:Chemistry; D004247:DNA; D006147:Guanine / Q000032:analysis; D007834:Lasers; D009690:Nucleic Acid Conformation; D011089:Polydeoxyribonucleotides; D012542:Scattering, Radiation; D013057:Spectrum Analysis
D011089:Polydeoxyribonucleotides; D006147:Guanine; D004247:DNA