TY - JOUR
T1 - Interstrand DNA Cross-Links Derived from Reaction of a 2-Aminopurine Residue with an Abasic Site
AU - Nejad, Maryam Imani
AU - Price, Nathan E.
AU - Haldar, Tuhin
AU - Lewis, Calvin
AU - Wang, Yinsheng
AU - Gates, Kent S.
N1 - Funding Information:
K.S.G. and Y.W. thank the National Institute of Environmental Health Science of the National Institutes of Health for support of this work (Grant ES 021007). NP was supported in part by an NRSA T32 institutional training grant (Grant ES018827).
Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/6/4
Y1 - 2019/6/4
N2 - Efficient methods for the site-specific installation of structurally defined interstrand cross-links in duplex DNA may be useful in a wide variety of fields. The work described here developed a high-yield synthesis of chemically stable interstrand cross-links resulting from a reductive amination reaction between an abasic site and the noncanonical nucleobase 2-aminopurine in duplex DNA. Results from footprinting, liquid chromatography-mass spectrometry, and stability studies support the formation of an N2-alkylamine attachment between the 2-aminopurine residue and the Ap site. The reaction performs best when the 2-aminopurine residue on the opposing strand is offset 1 nt to the 5′-side of the abasic site. The cross-link confers substantial resistance to thermal denaturation (melting). The cross-linking reaction is fast (complete in 4 h), employs only commercially available reagents, and can be used to generate cross-linked duplexes in sufficient quantities for biophysical, structural, and DNA repair studies.
AB - Efficient methods for the site-specific installation of structurally defined interstrand cross-links in duplex DNA may be useful in a wide variety of fields. The work described here developed a high-yield synthesis of chemically stable interstrand cross-links resulting from a reductive amination reaction between an abasic site and the noncanonical nucleobase 2-aminopurine in duplex DNA. Results from footprinting, liquid chromatography-mass spectrometry, and stability studies support the formation of an N2-alkylamine attachment between the 2-aminopurine residue and the Ap site. The reaction performs best when the 2-aminopurine residue on the opposing strand is offset 1 nt to the 5′-side of the abasic site. The cross-link confers substantial resistance to thermal denaturation (melting). The cross-linking reaction is fast (complete in 4 h), employs only commercially available reagents, and can be used to generate cross-linked duplexes in sufficient quantities for biophysical, structural, and DNA repair studies.
UR - http://www.scopus.com/inward/record.url?scp=85070117767&partnerID=8YFLogxK
U2 - 10.1021/acschembio.9b00208
DO - 10.1021/acschembio.9b00208
M3 - Article
C2 - 31259519
AN - SCOPUS:85070117767
SN - 1554-8929
VL - 14
SP - 1481
EP - 1489
JO - ACS Chemical Biology
JF - ACS Chemical Biology
IS - 7
ER -