Laser crosslinking of E. coli RNA polymerase and T7 DNA.
The first photochemical crosslinking of a protein to a nucleic acid using laser excitation is reported. A single, 120 mJ, 20 ns pulse at 248 nm crosslinks about 10% of bound E. coli RNA polymerase to T7 DNA under the conditions studied. The crosslinking yield depends on mercaptoethanol concentration, and is a linear function of laser intensity. The protein subunits crosslinked to DNA are beta, beta' and sigma.
D004279:DNA, Viral / Q000378:metabolism / Q000528:radiation effects*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism / Q000528:radiation effects*; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D007834:Lasers; D046911:Macromolecular Substances; D011485:Protein Binding; D013604:T-Phages / Q000378:metabolism*
D004279:DNA, Viral; D046911:Macromolecular Substances; D012321:DNA-Directed RNA Polymerases