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The structural effects nanobodies towards Gai

2024
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Poster Description

G protein coupled receptors (GPCRs) are responsible for converting extracellular messenger signals (ligands) to activation of metabolic pathways intracellularly. These signals are transduced using guanine nucleotide binding proteins (G proteins) that exchange GDP for GTP. G proteins are heterotrimers consisting of alpha ( ), beta (β), and gamma (γ) subunits. We have designed two nanobodies (NB), or antibody fragments, with high affinity for the G i subunit. Our research features two such nanobodies; NB9155 and NB9156, with the goal of understanding how G i:NB complex formation will affect protein stability and structure. As NB9155 and NB9156 have different binding sites for G i, we also investigate nanobody effects on guanine nucleotide exchange rates and potential chaperone activity. Our data will include confirmation of molecular weights using the Multi-Angle Light Scattering technique, monitoring nucleotide exchange rates using a Fluorescence Spectrometer, and comparison of crystal structures, determined by X-ray diffraction, for the G i:NB9155 and the G i:NB9156 crystal structures.

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