Methods to Monitor Ras Activation State

Kari Kopra; Harri Härmä

doi:10.1007/978-1-0716-1190-6_8

Methods to Monitor Ras Activation State

Methods Mol Biol. 2021:2262:137-167. doi: 10.1007/978-1-0716-1190-6_8.

Authors

Kari Kopra¹, Harri Härmä²

Affiliations

¹ Department of Chemistry, Chemistry of Drug Development, University of Turku, Turku, Finland. khkopr@utu.fi.
² Department of Chemistry, Chemistry of Drug Development, University of Turku, Turku, Finland.

PMID: 33977475
DOI: 10.1007/978-1-0716-1190-6_8

Abstract

Various biochemical methods have been introduced to detect and characterize small GTPases and Ras. Luminescence-based techniques cover most of the currently used methods, utilizing single- or multi-luminophore-conjugated molecules and external probes. Here we describe methods enabling Ras activity and activation state monitoring in vitro. This chapter focuses mainly on luminescence-based techniques.

Keywords: Environment-sensitive labels; Fluorescence polarization (FP); Förster resonance energy transfer (FRET); GTP- and GDP-specific antibodies; Quenching resonance energy transfer (QRET); Ras; Ras-binding domain (RBD); Small GTPases; Thermal stability assay (TSA); Time-resolved fluorescence resonance energy transfer (TR-FRET).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Fluorescence Resonance Energy Transfer / methods*
Guanosine Triphosphate / metabolism*
Humans
Luminescent Measurements / methods*
ras Proteins / metabolism*

Substances

Guanosine Triphosphate
ras Proteins