Crosslinking of diene-modified DNA with bis-maleimides

Abstract

The chemical crosslinking of modified nucleic acids via the Diels-Alder reaction is reported. For this purpose, 1,3-butadiene derived building blocks were incorporated into complementary oligodeoxynucleotides. Treatment of the obtained duplex with difunctional dienophiles results in the clean crosslinking of the two strands. Non-crosslinked adducts arising from a single Diels-Alder reaction of a maleimide to only one strand were not observed, indicating that the first reaction is the rate determining step of the overall process. Based on their thermal denaturation profiles, the crosslinked hybrids behave like two separate, hairpin-like structures, rather than like a single, continuous duplex.

Synthesis of misfolded glycoprotein dimers through native chemical ligation of a dimeric peptide thioester

“…This peptide was Scheme 1 Synthesis of monomeric misfolded M9-IL-8 analogs 5, 7-9. This peptide thioester 10 containing a Cys residue was reacted with 1,2-ethylene bismaleimide (11) 27,28 in 6 M Gn•HCl at pH 5.0 to yield the desired dimeric peptide-α-thioester 12 along with a monomeric peptide that had reacted with the bismaleimide linker (Fig. S7 †).…”

Novel strategies for the site-specific covalent labelling of nucleic acids

“…Later on crosslinking of complementary strands could be achieved using a bismaleinimide and this butadiene building block. 80 Recently, oligonucleotide-peptide conjugates were prepared using hexadiene 5 0 modified oligonucleotides and maleinimide modified peptides. 81 Over all, the Diels-Alder cycloaddition is a promising reaction for DNA conjugation.…”

Rapid chemical ligation of oligonucleotides by the Diels–Alder reaction

“…The process is slow (7 days) because the initial Diels-Alder reactions is not templated. 14 Diels-Alder chemistry has also been used to immobilise oligonucleotides on surfaces. 15…”