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Studies on the functional topography of <em> Escherichia coli</em> RNA polymerase.
- Source :
- European Journal of Biochemistry; 2/16/87, Vol. 163 Issue 1, p113-121, 9p
- Publication Year :
- 1987
-
Abstract
- RNA polymerase was treated in the presence of promoter-containing templates with 16 affinity reagents, derivatives on NMPs, NDPs and NTPs with reactive substituents at the terminal phosphate. This treatment was followed by addition of a pyrimidine [α-<superscript>32</superscript>P]NTP. Due to 'catalytic competence' of some of the residues of the affinity reagents bound covalently near the active center at the first stage, active-center-catalyzed synthesis of a phosphodiester bond occurred, and radioactive residues with the general formula -pNpN (where p = radioactive phosphate) appeared covalently attached to the enzyme. Such affinity labelling was super-selective because affinity reagent residues bound outside the active center were not elongated and thus remained non-radioactive. Labelling took place only when the combination of the reagent and [α-<superscript>32</superscript>P]NTP corresponded to the sequence of nucleotides of the promoter. With reagents having short 'arms', only the β subunit was labelled; the targets were His and/or Lys residues. With reagents having longer 'arms', the a subunit was also labelled. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 00142956
- Volume :
- 163
- Issue :
- 1
- Database :
- Complementary Index
- Journal :
- European Journal of Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 13746682
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1987.tb10743.x