The structure of the zinc sites of Escherichia coli DNA-dependent RNA polymerase

Felicia Y H Wu, Wei Jeu Huang, Robert B. Sinclair, Linda S Powers

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

X-ray absorption spectroscopy is ideally suited for the investigation of the electronic structure and the local environment (∼5 Å) of specific atoms in biomolecules. While the edge region provides information about the valence state of the absorbing atom, the chemical identity of neighboring atoms, and the coordination geometry, the extended x-ray absorption fine structure region contains information about the number and average distance of neighboring atoms and their relative disorder. The development of sensitive detection methods has allowed studies using near physiological concentrations (as low as ∼100 μM). RNA polymerase from Escherichia coli contains two zinc atoms: one tightly bound in the β′ subunit, the subunit that participates in template binding, and the other loosely bound in the β subunit, the subunit that participates in substrate binding. X-ray absorption studies of these zinc sites in the native protein and of the zinc site in the β′ subunit after removal of the zinc in the β subunit site by p-(hydroxymercuri)benzenesulfonate (Giedroc, D. P., and Coleman, J. E. (1986) Biochemistry 25, 4969-4978) indicate that both zinc sites have octahedral coordination. The zinc in the β′ subunit site has four sulfur ligands at an average distance of 2.36 ± 0.02 Å and two oxygen (or nitrogen) ligands at an average distance of 2.23 ± 0.02 Å. The β subunit zinc site has five sulfur ligands at an average distance of 2.38 ± 0.01 Å and one histidine nitrogen ligand at 2.14 ± 0.02 Å. These results are in general agreement with earlier biochemical and spectroscopic studies.

Original languageEnglish (US)
Pages (from-to)25560-25567
Number of pages8
JournalJournal of Biological Chemistry
Volume267
Issue number35
StatePublished - Dec 15 1992
Externally publishedYes

Fingerprint

DNA-Directed RNA Polymerases
Escherichia coli
Zinc
Atoms
Ligands
Sulfur
Nitrogen
X-Ray Absorption Spectroscopy
X-Rays
Biochemistry
X ray absorption spectroscopy
X ray absorption
Biomolecules
Histidine
Electronic structure
Oxygen
X rays
Geometry
Substrates
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

The structure of the zinc sites of Escherichia coli DNA-dependent RNA polymerase. / Wu, Felicia Y H; Huang, Wei Jeu; Sinclair, Robert B.; Powers, Linda S.

In: Journal of Biological Chemistry, Vol. 267, No. 35, 15.12.1992, p. 25560-25567.

Research output: Contribution to journalArticle

Wu, Felicia Y H ; Huang, Wei Jeu ; Sinclair, Robert B. ; Powers, Linda S. / The structure of the zinc sites of Escherichia coli DNA-dependent RNA polymerase. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 35. pp. 25560-25567.
@article{d58274acae3a4312bfdf95861c901f09,
title = "The structure of the zinc sites of Escherichia coli DNA-dependent RNA polymerase",
abstract = "X-ray absorption spectroscopy is ideally suited for the investigation of the electronic structure and the local environment (∼5 {\AA}) of specific atoms in biomolecules. While the edge region provides information about the valence state of the absorbing atom, the chemical identity of neighboring atoms, and the coordination geometry, the extended x-ray absorption fine structure region contains information about the number and average distance of neighboring atoms and their relative disorder. The development of sensitive detection methods has allowed studies using near physiological concentrations (as low as ∼100 μM). RNA polymerase from Escherichia coli contains two zinc atoms: one tightly bound in the β′ subunit, the subunit that participates in template binding, and the other loosely bound in the β subunit, the subunit that participates in substrate binding. X-ray absorption studies of these zinc sites in the native protein and of the zinc site in the β′ subunit after removal of the zinc in the β subunit site by p-(hydroxymercuri)benzenesulfonate (Giedroc, D. P., and Coleman, J. E. (1986) Biochemistry 25, 4969-4978) indicate that both zinc sites have octahedral coordination. The zinc in the β′ subunit site has four sulfur ligands at an average distance of 2.36 ± 0.02 {\AA} and two oxygen (or nitrogen) ligands at an average distance of 2.23 ± 0.02 {\AA}. The β subunit zinc site has five sulfur ligands at an average distance of 2.38 ± 0.01 {\AA} and one histidine nitrogen ligand at 2.14 ± 0.02 {\AA}. These results are in general agreement with earlier biochemical and spectroscopic studies.",
author = "Wu, {Felicia Y H} and Huang, {Wei Jeu} and Sinclair, {Robert B.} and Powers, {Linda S}",
year = "1992",
month = "12",
day = "15",
language = "English (US)",
volume = "267",
pages = "25560--25567",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "35",

}

TY - JOUR

T1 - The structure of the zinc sites of Escherichia coli DNA-dependent RNA polymerase

AU - Wu, Felicia Y H

AU - Huang, Wei Jeu

AU - Sinclair, Robert B.

AU - Powers, Linda S

PY - 1992/12/15

Y1 - 1992/12/15

N2 - X-ray absorption spectroscopy is ideally suited for the investigation of the electronic structure and the local environment (∼5 Å) of specific atoms in biomolecules. While the edge region provides information about the valence state of the absorbing atom, the chemical identity of neighboring atoms, and the coordination geometry, the extended x-ray absorption fine structure region contains information about the number and average distance of neighboring atoms and their relative disorder. The development of sensitive detection methods has allowed studies using near physiological concentrations (as low as ∼100 μM). RNA polymerase from Escherichia coli contains two zinc atoms: one tightly bound in the β′ subunit, the subunit that participates in template binding, and the other loosely bound in the β subunit, the subunit that participates in substrate binding. X-ray absorption studies of these zinc sites in the native protein and of the zinc site in the β′ subunit after removal of the zinc in the β subunit site by p-(hydroxymercuri)benzenesulfonate (Giedroc, D. P., and Coleman, J. E. (1986) Biochemistry 25, 4969-4978) indicate that both zinc sites have octahedral coordination. The zinc in the β′ subunit site has four sulfur ligands at an average distance of 2.36 ± 0.02 Å and two oxygen (or nitrogen) ligands at an average distance of 2.23 ± 0.02 Å. The β subunit zinc site has five sulfur ligands at an average distance of 2.38 ± 0.01 Å and one histidine nitrogen ligand at 2.14 ± 0.02 Å. These results are in general agreement with earlier biochemical and spectroscopic studies.

AB - X-ray absorption spectroscopy is ideally suited for the investigation of the electronic structure and the local environment (∼5 Å) of specific atoms in biomolecules. While the edge region provides information about the valence state of the absorbing atom, the chemical identity of neighboring atoms, and the coordination geometry, the extended x-ray absorption fine structure region contains information about the number and average distance of neighboring atoms and their relative disorder. The development of sensitive detection methods has allowed studies using near physiological concentrations (as low as ∼100 μM). RNA polymerase from Escherichia coli contains two zinc atoms: one tightly bound in the β′ subunit, the subunit that participates in template binding, and the other loosely bound in the β subunit, the subunit that participates in substrate binding. X-ray absorption studies of these zinc sites in the native protein and of the zinc site in the β′ subunit after removal of the zinc in the β subunit site by p-(hydroxymercuri)benzenesulfonate (Giedroc, D. P., and Coleman, J. E. (1986) Biochemistry 25, 4969-4978) indicate that both zinc sites have octahedral coordination. The zinc in the β′ subunit site has four sulfur ligands at an average distance of 2.36 ± 0.02 Å and two oxygen (or nitrogen) ligands at an average distance of 2.23 ± 0.02 Å. The β subunit zinc site has five sulfur ligands at an average distance of 2.38 ± 0.01 Å and one histidine nitrogen ligand at 2.14 ± 0.02 Å. These results are in general agreement with earlier biochemical and spectroscopic studies.

UR - http://www.scopus.com/inward/record.url?scp=0026487347&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026487347&partnerID=8YFLogxK

M3 - Article

VL - 267

SP - 25560

EP - 25567

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 35

ER -