TY  - INPR
A1  - Dopfer, Johannes
A1  - Schwalm, Martin P.
A1  - Knapp, Stefan
A1  - Rogov, Vladimir V.
T1  - Thermodynamic characterization of LC3/GABARAP ligands by isothermal titration calorimetry
T2  - bioRxiv
N2  - Isothermal titration calorimetry (ITC) is a widely used technique for the characterization of protein-protein and protein-ligand interactions. It provides information on the stoichiometry, affinity, and the thermodynamic driving forces of interactions. This chapter exemplifies the use of ITC to investigate interactions between human autophagy modifiers (LC3/GABARAP proteins) and their interaction partners, the LIR motif containing sequences. The purpose of this report is to present a detailed protocol for the production of LC3/GABARAP-interacting LIR peptides using E. coli expression systems. In addition, we outline the design of ITC experiments using the LC3/GABARAP:peptide interactions as an example. Comprehensive troubleshooting notes are provided to facilitate the adaptation of these protocols to different ligand-receptor systems. The methodology outlined for studying protein-ligand interactions will help to avoid common errors and misinterpretations of experimental results.
KW  - Isothermal Titration Calorimetry
KW  - Kd determination
KW  - autophagy
KW  - LC3
KW  - GABARAP
KW  - LIR motifs
Y1  - 2024
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/83030
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-830309
UR  - https://www.biorxiv.org/content/10.1101/2024.01.08.574610v1
IS  - 2024.01.08.574610 Version 1
PB  - bioRxiv
ER  -