Use of Site-Specifically Tethered Chemical Nucleases to Study Macromolecular Reactions
<p>During a complex macromolecular reaction multiple changes in molecular conformation and interactions with ligands may occur. X-ray crystallography may provide only a limited set of snapshots of these changes. Solution methods can augment such structural information to provide a more complet...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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BMC
2003-01-01
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| Series: | Biological Procedures Online |
| Subjects: | |
| Online Access: | http://www.biologicalprocedures.com/bpo/arts/1/49/m49.htm |
| Summary: | <p>During a complex macromolecular reaction multiple changes in molecular conformation and interactions with ligands may occur. X-ray crystallography may provide only a limited set of snapshots of these changes. Solution methods can augment such structural information to provide a more complete picture of a macromolecular reaction. We analyzed the changes in protein conformation and protein:nucleic acid interactions which occur during transcription initiation by using a chemical nuclease tethered to cysteines introduced site-specifically into the RNA polymerase of bacteriophage T7 (T7 RNAP). Changes in cleavage patterns as the polymerase steps through transcription reveal a series of structural transitions which mediate transcription initiation. Cleavage by tethered chemical nucleases is seen to be a powerful method for revealing the conformational dynamics of macromolecular reactions, and has certain advantages over cross-linking or energy transfer approaches. |
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| ISSN: | 1480-9222 |