Interaction of sigma(70) with Escherichia coli RNA polymerase core enzyme studied by surface plasmon resonance

A L Ferguson; A D Hughes; U Tufail; C G Baumann; D J Scott; J G Hoggett

Interaction of sigma(70) with Escherichia coli RNA polymerase core enzyme studied by surface plasmon resonance

A L Ferguson, A D Hughes, U Tufail, C G Baumann, D J Scott, J G Hoggett

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

The interaction between the core form of bacterial RNA polymerases and a factors is essential for specific promoter recognition, and for coordinating the expression of different sets of genes in response to varying cellular needs. The interaction between Escherichia coli core RNA polymerase and sigma(70) has been investigated by surface plasmon resonance. The His-tagged form of sigma(70) factor was immobilised on a Ni2+-NTA chip for monitoring its interaction with care polymerase, The binding constant for the interaction was found to be 1.9 x 10(-7) M, and the dissociation rate constant for release of a from core, in the absence of DNA or transcription, was 4 x 10(-3) s(-1), corresponding to a half-life of about 200 s, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

Original language	English
Pages (from-to)	281-284
Number of pages	4
Journal	FEBS Letters
Volume	481
Issue number	3
Publication status	Published - 22 Sept 2000

Keywords

surface plasmon resonance
RNA polymerase
sigma factors
Escherichia coli
PURIFICATION
SUBUNIT

Cite this

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title = "Interaction of sigma(70) with Escherichia coli RNA polymerase core enzyme studied by surface plasmon resonance",

abstract = "The interaction between the core form of bacterial RNA polymerases and a factors is essential for specific promoter recognition, and for coordinating the expression of different sets of genes in response to varying cellular needs. The interaction between Escherichia coli core RNA polymerase and sigma(70) has been investigated by surface plasmon resonance. The His-tagged form of sigma(70) factor was immobilised on a Ni2+-NTA chip for monitoring its interaction with care polymerase, The binding constant for the interaction was found to be 1.9 x 10(-7) M, and the dissociation rate constant for release of a from core, in the absence of DNA or transcription, was 4 x 10(-3) s(-1), corresponding to a half-life of about 200 s, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.",

keywords = "surface plasmon resonance, RNA polymerase, sigma factors, Escherichia coli, PURIFICATION, SUBUNIT",

author = "Ferguson, {A L} and Hughes, {A D} and U Tufail and Baumann, {C G} and Scott, {D J} and Hoggett, {J G}",

year = "2000",

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journal = "FEBS Letters",

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TY - JOUR

T1 - Interaction of sigma(70) with Escherichia coli RNA polymerase core enzyme studied by surface plasmon resonance

AU - Ferguson, A L

AU - Hughes, A D

AU - Tufail, U

AU - Baumann, C G

AU - Scott, D J

AU - Hoggett, J G

PY - 2000/9/22

Y1 - 2000/9/22

N2 - The interaction between the core form of bacterial RNA polymerases and a factors is essential for specific promoter recognition, and for coordinating the expression of different sets of genes in response to varying cellular needs. The interaction between Escherichia coli core RNA polymerase and sigma(70) has been investigated by surface plasmon resonance. The His-tagged form of sigma(70) factor was immobilised on a Ni2+-NTA chip for monitoring its interaction with care polymerase, The binding constant for the interaction was found to be 1.9 x 10(-7) M, and the dissociation rate constant for release of a from core, in the absence of DNA or transcription, was 4 x 10(-3) s(-1), corresponding to a half-life of about 200 s, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

AB - The interaction between the core form of bacterial RNA polymerases and a factors is essential for specific promoter recognition, and for coordinating the expression of different sets of genes in response to varying cellular needs. The interaction between Escherichia coli core RNA polymerase and sigma(70) has been investigated by surface plasmon resonance. The His-tagged form of sigma(70) factor was immobilised on a Ni2+-NTA chip for monitoring its interaction with care polymerase, The binding constant for the interaction was found to be 1.9 x 10(-7) M, and the dissociation rate constant for release of a from core, in the absence of DNA or transcription, was 4 x 10(-3) s(-1), corresponding to a half-life of about 200 s, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

KW - surface plasmon resonance

KW - RNA polymerase

KW - sigma factors

KW - Escherichia coli

KW - PURIFICATION

KW - SUBUNIT

M3 - Article

SN - 0014-5793

VL - 481

SP - 281

EP - 284

JO - FEBS Letters

JF - FEBS Letters

IS - 3

ER -