TY - GEN
T1 - Conversion of helix-turn-helix motif sequence-specific DNA binding proteins into site-specific DNA cleavage agents
AU - Ebright, Richard H.
AU - Ebright, Yon W.
AU - Pendergrast, P. Shannon
AU - Gunasekera, Angelo
PY - 1990
Y1 - 1990
N2 - An approach is presented to construct novel, semisynthetic site-specific DNA cleavage agents that have DNA recognition sites longer than those of the type II restriction endonucleases. To demonstrate the approach, a prototype was constructed in which the chelator 1,10-phenanthroline is incorporated at amino acid 10 of the helix-turn-helix motif of the Escherichia coli catabolite gene activator protein: [(N-acetyl-5-amino-1,10-phenanthroline)-Cys178]CAP. The prototype binds to a 22 base pair DNA recognition site with Kobs = 1 × 108 M-1. In the presence of Cu(II) and reducing agent, the prototype cleaves DNA at four adjacent nucleotides on each DNA strand within the DNA recognition site. The DNA cleavage reaction using 40 base pair and 7164 base pair DNA substrates is demonstrated.
AB - An approach is presented to construct novel, semisynthetic site-specific DNA cleavage agents that have DNA recognition sites longer than those of the type II restriction endonucleases. To demonstrate the approach, a prototype was constructed in which the chelator 1,10-phenanthroline is incorporated at amino acid 10 of the helix-turn-helix motif of the Escherichia coli catabolite gene activator protein: [(N-acetyl-5-amino-1,10-phenanthroline)-Cys178]CAP. The prototype binds to a 22 base pair DNA recognition site with Kobs = 1 × 108 M-1. In the presence of Cu(II) and reducing agent, the prototype cleaves DNA at four adjacent nucleotides on each DNA strand within the DNA recognition site. The DNA cleavage reaction using 40 base pair and 7164 base pair DNA substrates is demonstrated.
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M3 - Conference contribution
AN - SCOPUS:0025557801
SN - 0879425598
T3 - Proceedings of the Annual Conference on Engineering in Medicine and Biology
SP - 1616
BT - Biomedical Engineering Perspectives
PB - Publ by IEEE
T2 - Proceedings of the 12th Annual International Conference of the IEEE Engineering in Medicine and Biology Society
Y2 - 1 November 1990 through 4 November 1990
ER -