Using Surface Plasmon Resonance to Study SH2 Domain-Peptide Interactions

Gabrielle M Watson; Menachem J Gunzburg; Jacqueline A Wilce

doi:10.1007/978-1-0716-3393-9_10

Using Surface Plasmon Resonance to Study SH2 Domain-Peptide Interactions

Methods Mol Biol. 2023:2705:199-210. doi: 10.1007/978-1-0716-3393-9_10.

Authors

Gabrielle M Watson¹, Menachem J Gunzburg², Jacqueline A Wilce³

Affiliations

¹ The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
² Monash Institute of Pharmaceutical Sciences, Monash University, Melbourne, VIC, Australia.
³ Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia. jackie.wilce@monash.edu.

PMID: 37668975
DOI: 10.1007/978-1-0716-3393-9_10

Abstract

Biosensor measurement using surface plasmon resonance enables precise evaluation of peptide-protein interactions. It is a sensitive technique that provides kinetic and affinity data with very little sample and without the need for analyte labels. Here, we describe its application for the analysis of peptide interactions with the Grb7-SH2 domain prepared with a GST-tag for tethering to the chip surface. This has been successfully and reliably used for direct comparison of a range of peptides under different solution conditions as well as direct comparison of peptides flowed over different related SH2 domains in real time. We have used the BIAcore system and describe both the methodology for data collection and analysis, with principles also applicable to other biosensor platforms.

Keywords: GST fusion protein expression; Growth receptor-bound protein 7; Sensor chip preparation; Steady-state analysis; Surface plasmon resonance.

MeSH terms

Data Collection
Kinetics
Peptides
Surface Plasmon Resonance*
src Homology Domains*

Substances

Peptides