Maiti, Atanu ; Roy, Siddhartha (2005) Switching DNA-binding specificity by unnatural amino acid substitution Nucleic Acids Research, 33 (18). pp. 5896-5903. ISSN 0305-1048
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Official URL: http://nar.oxfordjournals.org/content/33/18/5896.a...
Related URL: http://dx.doi.org/10.1093/nar/gki899
Abstract
The specificity of protein-nucleic acid recognition is believed to originate largely from hydrogen bonding between protein polar atoms, primarily side-chain and polar atoms of nucleic acid bases. One way to design new nucleic acid binding proteins of novel specificity is by structure-guided alterations of the hydrogen bonding patterns of a nucleic acid-protein complex. We have used cI repressor of bacteriophage λ as a model system. In the λ-repressor-DNA complex, the ε-NH2 group (hydrogen bond donor) of lysine-4 of λ-repressor forms hydrogen bonds with the amide carbonyl atom of asparagine-55 (acceptor) and the O6 (acceptor) of CG6 of operator site OL1. Substitution of lysine-4 (two donors) by iso-steric S-(2-hydroxyethyl)-cysteine (one donor and one acceptor), by site-directed mutagenesis and chemical modification, leads to switch of binding specificity of λ-repressor from C:G to T:A at position 6 of OL1. This suggests that unnatural amino acid substitutions could be a simple way of generating nucleic acid binding proteins of altered specificity.
Item Type: | Article |
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Source: | Copyright of this article belongs to Oxford University Press. |
ID Code: | 43155 |
Deposited On: | 10 Jun 2011 06:25 |
Last Modified: | 18 May 2016 00:14 |
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