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 sigma 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 core polymerase. The binding constant for the interaction was found to be 1.9x10(-7) M, and the dissociation rate constant for release of sigma from core, in the absence of DNA or transcription, was 4x10(-3) s(-1), corresponding to a half-life of about 200 s.

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

Keywords

Bacterial Proteins
Binding, Competitive
DNA-Directed RNA Polymerases
Enzyme Stability
Escherichia coli
Half-Life
Histidine
Holoenzymes
Kinetics
Nickel
Nitrilotriacetic Acid
Organometallic Compounds
Protein Binding
Sigma Factor
Surface Plasmon Resonance

Cite this

@article{55ef4e990d4d4de995c46a5014b50c91,

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 sigma 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 core polymerase. The binding constant for the interaction was found to be 1.9x10(-7) M, and the dissociation rate constant for release of sigma from core, in the absence of DNA or transcription, was 4x10(-3) s(-1), corresponding to a half-life of about 200 s.",

keywords = "Bacterial Proteins, Binding, Competitive, DNA-Directed RNA Polymerases, Enzyme Stability, Escherichia coli, Half-Life, Histidine, Holoenzymes, Kinetics, Nickel, Nitrilotriacetic Acid, Organometallic Compounds, Protein Binding, Sigma Factor, Surface Plasmon Resonance",

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",

language = "English",

volume = "481",

pages = "281--4",

journal = "FEBS Letters",

issn = "0014-5793",

publisher = "Wiley-Blackwell",

number = "3",

}

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

Y1 - 2000

N2 - The interaction between the core form of bacterial RNA polymerases and sigma 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 core polymerase. The binding constant for the interaction was found to be 1.9x10(-7) M, and the dissociation rate constant for release of sigma from core, in the absence of DNA or transcription, was 4x10(-3) s(-1), corresponding to a half-life of about 200 s.

AB - The interaction between the core form of bacterial RNA polymerases and sigma 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 core polymerase. The binding constant for the interaction was found to be 1.9x10(-7) M, and the dissociation rate constant for release of sigma from core, in the absence of DNA or transcription, was 4x10(-3) s(-1), corresponding to a half-life of about 200 s.

KW - Bacterial Proteins

KW - Binding, Competitive

KW - DNA-Directed RNA Polymerases

KW - Enzyme Stability

KW - Escherichia coli

KW - Half-Life

KW - Histidine

KW - Holoenzymes

KW - Kinetics

KW - Nickel

KW - Nitrilotriacetic Acid

KW - Organometallic Compounds

KW - Protein Binding

KW - Sigma Factor

KW - Surface Plasmon Resonance

M3 - Article

C2 - 11007979

SN - 0014-5793

VL - 481

SP - 281

EP - 284

JO - FEBS Letters

JF - FEBS Letters

IS - 3

ER -