Microscale Thermophoresis (MST ) as a Tool for Measuring Dynamin Superfamily Protein (DSP)-Lipid Interactions

Nikhil Bharambe; Rajesh Ramachandran

doi:10.1007/978-1-0716-0676-6_7

Microscale Thermophoresis (MST ) as a Tool for Measuring Dynamin Superfamily Protein (DSP)-Lipid Interactions

Methods Mol Biol. 2020:2159:85-92. doi: 10.1007/978-1-0716-0676-6_7.

Authors

Nikhil Bharambe¹, Rajesh Ramachandran^{2

3}

Affiliations

¹ Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
² Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, Cleveland, OH, USA. rxr275@case.edu.
³ Cleveland Center for Membrane and Structural Biology, Case Western Reserve University School of Medicine, Cleveland, OH, USA. rxr275@case.edu.

PMID: 32529365
DOI: 10.1007/978-1-0716-0676-6_7

Abstract

Microscale thermophoresis (MST ) is a robust new fluorescence-based technology that enables measurement of biomolecular interactions and binding affinities (K_D). MST is an immobilization-free alternative to surface plasmon resonance (SPR ) and is cost-effective relative to isothermal titration calorimetry (ITC ). In this chapter, using Drp1 as an example, we demonstrate for the first time, the application of MST to the determination of DSP-lipid interactions and the accurate measurement of K_D under physiologically relevant solution conditions.

Keywords: Drp1 variable domain; Dynamin-related protein 1; Equilibrium dissociation constant; Microscale thermophoresis; Protein–lipid interactions.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Calorimetry* / methods
Data Analysis
Dynamins / chemistry*
Dynamins / metabolism
Kinetics
Lipids / chemistry*
Protein Binding
Surface Plasmon Resonance* / methods
Temperature

Substances

Lipids
Dynamins

Grants and funding

R01 GM121583/GM/NIGMS NIH HHS/United States