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Published on: June 3, 2021
Structural basis of latrophilin-FLRT interaction
Verity A Jackson1, Daniel del Toro2, Maria Carrasquero1
1Department of Biochemistry, Oxford University, South Parks Road, Oxford OX1 3QU, UK.
Abstract:
Latrophilins, receptors for spider venom α-latrotoxin, are adhesion type G-protein-coupled receptors with emerging functions in synapse development. The N-terminal region binds the endogenous cell adhesion molecule FLRT, a major regulator of cortical and synapse development. We present crystallographic data for the mouse Latrophilin3 lectin and olfactomedin-like (Olf) domains, thereby revealing the Olf β-propeller fold and conserved calcium-binding site. We locate the FLRT-Latrophilin binding surfaces by a combination of sequence conservation analysis, point mutagenesis, and surface plasmon resonance experiments. In stripe assays, we show that wild-type Latrophilin3 and its high-affinity interactor FLRT2, but not the binding-impaired mutants we generated, promote HeLa cell adhesion. In contrast, cortical neurons expressing endogenous FLRTs are repelled by wild-type Latrophilin3 and not by the binding-impaired mutant. Taken together, we present molecular level insights into Latrophilin structure, its FLRT-binding mechanism, and a role for Latrophilin and FLRT that goes beyond a simply adhesive interaction.
Insights
Latrophilins are G-protein-coupled receptors involved in synapse development. This study reveals their structure, how they bind FLRT, and a complex role in cell adhesion beyond simple attraction.
Area of Science:
- Neuroscience
- Molecular Biology
- Structural Biology
Background:
- Latrophilins are G-protein-coupled receptors crucial for synapse development.
- They interact with FLRT, a key regulator of cortical and synapse formation.
Purpose of the Study:
- To elucidate the molecular structure of Latrophilin3 domains.
- To identify the binding mechanism between Latrophilin and FLRT.
- To investigate the functional role of Latrophilin-FLRT interactions in cell adhesion.
Main Methods:
- X-ray crystallography to determine the structure of Latrophilin3 lectin and olfactomedin-like (Olf) domains.
- Sequence conservation analysis, point mutagenesis, and surface plasmon resonance to map binding interfaces.
- Stripe assays using HeLa cells and cortical neurons to assess cell adhesion.
Main Results:
- The Olf domain exhibits a β-propeller fold with a conserved calcium-binding site.
- Key binding surfaces between Latrophilin and FLRT were identified.
- Wild-type Latrophilin3 promoted HeLa cell adhesion, while binding-impaired mutants did not.
- Cortical neurons were repelled by wild-type Latrophilin3, unlike the mutants.
Conclusions:
- Molecular insights into Latrophilin structure and its interaction with FLRT were provided.
- The study demonstrates a complex role for Latrophilin and FLRT in cell adhesion, involving both attraction and repulsion.
- Findings advance understanding of synapse development regulation by adhesion molecules.
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