title: γ-Hydroxy-1,N2-propano-2'-deoxyguanosine DNA adduct conjugates the N-terminal amine of the KWKK peptide via a carbinolamine linkage.
abstract: The γ-hydroxy-1,N(2)-propano-2'-deoxyguanosine adduct (γ-OH-PdG) was introduced into 5'-d(GCTAGCXAGTCC)-3'·5'-d(GGACTCGCTAGC)-3' (X = γ-OH-PdG). In the presence of excess peptide KWKK, (13)C isotope-edited NMR revealed the formation of two spectroscopically distinct DNA-KWKK conjugates. These involved the reaction of the KWKK N-terminal amino group with the N(2)-dG propylaldehyde tautomer of the γ-OH-PdG lesion. The guanine N1 base imino resonance at the site of conjugation was observed in isotope-edited (15)N NMR experiments, suggesting that the conjugated guanine was inserted into the duplex and that the guanine imino proton was protected from exchange with water. The conjugates could be reduced in the presence of NaCNBH(3), suggesting that they existed, in part, as imine (Schiff base) linkages. However, (13)C isotope-edited NMR failed to detect the imine linkages, suggesting that these KWKK conjugates existed predominantly as diastereomeric carbinolamines, in equilibrium with trace amounts of the imines. The structures of the diastereomeric DNA-KWKK conjugates were predicted from potential energy minimization of model structures derived from the refined structure of the fully reduced cross-link [ Huang, H., Kozekov, I. D., Kozekova, A., Rizzo, C. J., McCullough, A., Lloyd, R. S., and Stone, M. P. ( 2010 ) Biochemistry , 49 , 6155 -6164 ]. Molecular dynamics calculations carried out in explicit solvent suggested that the conjugate bearing the S-carbinolamine linkage was the major species due to its potential for intramolecular hydrogen bonding. These carbinolamine DNA-KWKK conjugates thermally stabilized duplex DNA. However, the DNA-KWKK conjugates were chemically reversible and dissociated when the DNA was denatured. In this 5'-CpX-3' sequence, the DNA-KWKK conjugates slowly converted to interstrand N(2)-dG:N(2)-dG DNA cross-links and ring-opened γ-OH-PdG derivatives over a period of weeks.
authors.affiliation: Department of Chemistry, Center in Molecular Toxicology, Center for Structural Biology, Vanderbilt University, Nashville, Tennessee 37235, USA.
authors.forename: Hai
authors.initials: H
authors.lastname: Huang
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.forename: Markus W
authors.initials: MW
authors.lastname: Voehler
authors.forename: Albena
authors.initials: A
authors.lastname: Kozekova
authors.forename: Carmelo J
authors.initials: CJ
authors.lastname: Rizzo
authors.forename: Amanda K
authors.initials: AK
authors.lastname: McCullough
authors.forename: R Stephen
authors.initials: RS
authors.lastname: Lloyd
authors.forename: Michael P
authors.initials: MP
authors.lastname: Stone
mesh_terms: D000171:Acrolein / Q000737:chemistry / Q000633:toxicity; D000588:Amines / Q000737:chemistry*; D000595:Amino Acid Sequence; D018736:DNA Adducts / Q000737:chemistry*; D003849:Deoxyguanosine / Q000031:analogs & derivatives*; D006860:Hydrogen Bonding; D007097:Imines / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D056004:Molecular Dynamics Simulation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D044366:Transition Temperature
chemical_list: D000588:Amines; D018736:DNA Adducts; D007097:Imines; D009838:Oligodeoxyribonucleotides; D010455:Peptides; D000171:Acrolein; D003849:Deoxyguanosine