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Switching DNA-binding specificity by unnatural amino acid substitution
| Content Provider | Semantic Scholar |
|---|---|
| Author | Maiti, Atanu Roy, Siddhartha |
| Copyright Year | 2005 |
| 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 lambda as a model system. In the lambda-repressor-DNA complex, the epsilon-NH(2) group (hydrogen bond donor) of lysine-4 of lambda-repressor forms hydrogen bonds with the amide carbonyl atom of asparagine-55 (acceptor) and the O6 (acceptor) of CG6 of operator site O(L)1. 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 lambda-repressor from C:G to T:A at position 6 of O(L)1. This suggests that unnatural amino acid substitutions could be a simple way of generating nucleic acid binding proteins of altered specificity. |
| Starting Page | 5896 |
| Ending Page | 5903 |
| Page Count | 8 |
| File Format | PDF HTM / HTML |
| PubMed reference number | 16224104 |
| Volume Number | 33 |
| Journal | Nucleic acids research |
| Alternate Webpage(s) | http://ftp.ncbi.nlm.nih.gov/pub/pmc/b7/54/gki899.PMC1258173.pdf |
| Alternate Webpage(s) | http://repository.ias.ac.in/43155/1/30-Pub.pdf |
| Alternate Webpage(s) | https://oup.silverchair-cdn.com/oup/backfile/Content_public/Journal/nar/33/18/10.1093_nar_gki899/2/gki899.pdf?Expires=1492920175&Key-Pair-Id=APKAIUCZBIA4LVPAVW3Q&Signature=ezGusYKO7Aa9IRZpEyyITnM8sVToB-TMoQGVb272bff~Zglzh8I7EgOxP85ol8gtpDcqSMVVa4lJFd~03yxaLpdxB05RPKd1gye6OX8Cj8u9MS4GJ5B4v480PhvRqdQwGZ5udY~uVPF0qwgEHHIAL-CX3qdCyldWUNKvpdRCzTgAJCeKM9xWcK7iA75dIS7Yr8bdhes6631Fg6abQStBIYlkAfO0inJbjJQ9H-tF5GUsWGSwhggpH7u9Pj9EEFby0csMlL3ygUSo82lAF0C-66KMMotxl2aAmZyuMPT6qLUvUdBX9ov50W2uwZvUrlf3EssX2c9Chi0CHCb~J7~rwg__ |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Amino Acid Substitution Amino Acids Bacteriophages Base Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone Cell Nucleus FRRS1L gene Hydrogen Bonding Isoproterenol Nucleic Acid Binding Nucleic Acids Pyschological Bonding Structural Modifier Transcription Repressor/Corepressor |
| Content Type | Text |
| Resource Type | Article |
