A rapid mixing-photocrosslinking technique to study the dynamics of nucleic acid-protein interactions.
A rapid mixing-photocrosslinking technique has been developed to investigate the kinetics of protein-nucleic acid interactions. With this technique, binding of nucleic acid to protein is first synchronized by rapid mixing in a stopped-flow apparatus. The intermediates formed at different stages of the binding process are then "frozen" by photocrosslinking with a 10-microseconds uv light pulse at various times after mixing. By analyzing structural changes of these intermediates as a function of time, one can obtain the information concerning the dynamic aspects of the interaction. This technique may also be applied to other macromolecular interactions in biological systems.
D004247:DNA / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D009696:Nucleic Acids / Q000378:metabolism*; D010777:Photochemistry / Q000295:instrumentation* / Q000379:methods; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D014466:Ultraviolet Rays
D009696:Nucleic Acids; D011506:Proteins; D004247:DNA; D012321:DNA-Directed RNA Polymerases