High-Throughput Protein-Nucleic Acid Interaction Assay Based on
Protein-Induced Fluorescence Enhancement

C 2021

High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement

FULNEČEK, Jaroslav and Karel ŘÍHA

Basic information

Original name

High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement

Authors

FULNEČEK, Jaroslav (203 Czech Republic, belonging to the institution) and Karel ŘÍHA (203 Czech Republic, guarantor, belonging to the institution)

Edition

TOTOWA, RNA REMODELING PROTEINS, p. 109-117, 9 pp. 2 EDITION, 2021

Publisher

HUMANA PRESS INC

Other information

Language

English

Type of outcome

Kapitola resp. kapitoly v odborné knize

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Publication form

electronic version available online

References:

URL

RIV identification code

RIV/00216224:14740/21:00119726

Organization unit

Central European Institute of Technology

ISBN

978-1-0716-0934-7

DOI

http://dx.doi.org/10.1007/978-1-0716-0935-4_7

UT WoS

000683504600008

Keywords in English

Protein-induced fluorescence enhancement (PIFE); Cy3; RNA; DNA; Protein; Binding assay

Abstract

V originále

Molecular processes involved in gene expression encompass multitudes of interactions between proteins and nucleic acids. Quantitative description of these interactions is crucial for delineating the mechanisms governing transcription, genome duplication, and translation. Here we describe a detailed protocol for the quantitative analysis of protein-nucleic acid interactions based on protein-induced fluorescence enhancement (PIFE). While PIFE has mainly been used in single-molecule studies, we modified its application for bulk measurement of protein-nucleic acid interactions in microwell plates using standard fluorescent plate readers. The microwell plate PIFE assay (mwPIFE) is simple, does not require laborious protein labeling, and is high throughput. These properties predispose mwPIFE to become a method of choice for routine applications that require multiple parallel measurements such as buffer optimization, competition experiments, or screening chemical libraries for binding modulators.

Links

EF15_003/0000479, research and development project

Name: Regulace rostlinné meiózy

GA19-21961S, research and development project

Name: Chemická modulace aktivity Ku komplexu a využití v editaci genomů.

Investor: Czech Science Foundation, Chemical modulation of Ku activity and its use in genome editing

Citovat

FULNEČEK, Jaroslav and Karel ŘÍHA. High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement. Online. In RNA REMODELING PROTEINS. TOTOWA: HUMANA PRESS INC, 2021, p. 109-117. 2 EDITION. ISBN 978-1-0716-0934-7. Available from: https://dx.doi.org/10.1007/978-1-0716-0935-4_7.

@inbook{1844939,
   author = {Fulneček, Jaroslav and Říha, Karel},
   address = {TOTOWA},
   booktitle = {RNA REMODELING PROTEINS},
   doi = {http://dx.doi.org/10.1007/978-1-0716-0935-4_7},
   keywords = {Protein-induced fluorescence enhancement (PIFE); Cy3; RNA; DNA; Protein; Binding assay},
   howpublished = {elektronická verze "online"},
   language = {eng},
   location = {TOTOWA},
   isbn = {978-1-0716-0934-7},
   pages = {109-117},
   publisher = {HUMANA PRESS INC},
   title = {High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement},
   url = {https://link.springer.com/protocol/10.1007/978-1-0716-0935-4_7#citeas},
   year = {2021}
}

TY  - CHAP
ID  - 1844939
AU  - Fulneček, Jaroslav - Říha, Karel
PY  - 2021
TI  - High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement
VL  - 2 EDITION
PB  - HUMANA PRESS INC
CY  - TOTOWA
SN  - 9781071609347
KW  - Protein-induced fluorescence enhancement (PIFE)
KW  - Cy3
KW  - RNA
KW  - DNA
KW  - Protein
KW  - Binding assay
UR  - https://link.springer.com/protocol/10.1007/978-1-0716-0935-4_7#citeas
N2  - Molecular processes involved in gene expression encompass multitudes of interactions between proteins and nucleic acids. Quantitative description of these interactions is crucial for delineating the mechanisms governing transcription, genome duplication, and translation. Here we describe a detailed protocol for the quantitative analysis of protein-nucleic acid interactions based on protein-induced fluorescence enhancement (PIFE). While PIFE has mainly been used in single-molecule studies, we modified its application for bulk measurement of protein-nucleic acid interactions in microwell plates using standard fluorescent plate readers. The microwell plate PIFE assay (mwPIFE) is simple, does not require laborious protein labeling, and is high throughput. These properties predispose mwPIFE to become a method of choice for routine applications that require multiple parallel measurements such as buffer optimization, competition experiments, or screening chemical libraries for binding modulators.
ER  -

FULNEČEK, Jaroslav and Karel ŘÍHA. High-Throughput Protein-Nucleic Acid Interaction Assay Based on Protein-Induced Fluorescence Enhancement. Online. In \textit{RNA REMODELING PROTEINS}. TOTOWA: HUMANA PRESS INC, 2021, p.~109-117. 2 EDITION. ISBN~978-1-0716-0934-7. Available from: https://dx.doi.org/10.1007/978-1-0716-0935-4\_{}7.

Detailed Information on Publication Record