A. Ishihama, Molecular anatomy of RNA polymerase using protein-conjugated metal probes with nuclease and protease activities, CHEM COMMUN, (13), 2000, pp. 1091-1094
Iron (S)-1-(p-bromoacetamidobenzyl)ethylenediaminetetraacetate (FeBABE) wit
h the sequence-non-specific cleavage activity of nucleic acids and proteins
was conjugated to protein Cys residues, and used for mapping the contact s
ites of both the alpha-subunit carboxy-terminal domain of Escherichia coli
RNA polymerase on promoter UP elements and the sigma(70) and sigma(38) subu
nits on the respective promoters. The same chemical nuclease was also used
as a chemical protease for mapping the subunit-subunit contact sites within
the RNA polymerase. By using 2-iminothiolane as a linker, FeBABE could be
conjugated to protein Lys residues and successfully used for mapping the co
ntact surfaces of some E. coli transcription factors on the RNA polymerase
holoenzyme.