Design, synthesis, and analysis of disulfide cross-linked DNA duplexes

Scott E. Osborne, Jens Völker, Shawn Y. Stevens, Kenneth J. Breslauer, Gary D. Glick

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

The design, synthesis, and analysis of analogs of d(CGCGAATTCGCG)2 possessing one or two intrahelical disulfide cross-links is reported. The cross-linked oligomers were prepared by first synthesizing duplexes where the 3'- and 5'-terminal bases of the parent sequence were replaced with N3-thioethylthymidine. Following deprotection and purification, air oxidation afforded the desired cross-linked constructs in high yield. Analysis of both the oxidized (disulfide cross-linked) and reduced (thiol modified) duplexes by UV, circular dichioism, and NMR spectroscopies along with susceptibility to EcoRI cleavage indicates that the modifications are not structurally perturbing. Optical thermal denaturation and differential scanning calorimetry measurements suggest that introducing disulfide cross-link(s) into d(CGCGAATTCGCG)2 does, however, cause two fundamental changes. First, the cross-link(s) increase the thermal stability of the modified duplexes by changing the molecularity of denaturation without an increase in enthalpy. Second, the disulfide cross-link traps one of the conformations of the conformationally heterogeneous parent molecule resulting in a conformationally homogeneous system. Both of these features are themselves unique and will be important for further applications of disulfide cross-linked oligomers such as these in studies of nucleic acid structure and function.

Original languageEnglish (US)
Pages (from-to)11993-12003
Number of pages11
JournalJournal of the American Chemical Society
Volume118
Issue number48
DOIs
StatePublished - 1996

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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