PMID:9477962
| Citation |
Miyake, R, Murakami, K, Owens, JT, Greiner, DP, Ozoline, ON, Ishihama, A and Meares, CF (1998) Dimeric association of Escherichia coli RNA polymerase alpha subunits, studied by cleavage of single-cysteine alpha subunits conjugated to iron-(S)-1-[p-(bromoacetamido)benzyl]ethylenediaminetetraacetate. Biochemistry 37:1344-9 |
|---|---|
| Abstract |
Proximity relationships between the two associated monomers of the Escherichia coli RNA polymerase alpha subunit were studied using a set of four mutant alpha subunits, each with a single Cys residue at one of the naturally occurring positions (54, 131, 176, and 269). These mutant alpha subunits were conjugated with the cutting reagent iron-(S)-1-[p-(bromoacetamido)benzyl]ethylenediaminetetraacetate (Fe-BABE), and the peptide backbone was cleaved at locations near the modified Cys. Analysis of the cleavage sites identified segments within approximately 12 A of the conjugation site. These results show that, for intermolecular cutting, segments of the subunit assembly domain (N-terminal domain) of one subunit and the linker region between N- and C-terminal domains of the other subunit are near each other, and the N-terminal domains of both subunits are in close proximity to one another. Intramolecular cutting however, was observed only within an individual N- or C-terminal domain. |
| Links |
PubMed Online version:10.1021/bi9723313 |
| Keywords |
Alkaline Phosphatase; Biotinylation; Cysteine/genetics; Cysteine/metabolism; DNA-Directed RNA Polymerases/chemical synthesis; DNA-Directed RNA Polymerases/genetics; DNA-Directed RNA Polymerases/metabolism; Dimerization; Edetic Acid/analogs & derivatives; Edetic Acid/metabolism; Escherichia coli/enzymology; Escherichia coli/genetics; Hydrolysis; Iron Chelating Agents/metabolism; Models, Molecular; Mutagenesis, Site-Directed; Organometallic Compounds/metabolism; Protein Folding; Streptavidin |
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