Investigation of the Lipid Transfer ProteinVPS13D at the Golgi Apparatus

Summary

This project aims to investigate the precise localization, recruitment, and function of the lipid transfer protein VPS13D at the Golgi apparatus, particularly its role in maintaining the trans-Golgi network (TGN) and its implications in neurodegenerative diseases.

What they want

The project involves three main aims: (1) elucidating the precise localization of VPS13D within the Golgi complex using techniques like nocodazole ministacks, superresolution, and immune-electron microscopy on an endogenously tagged VPS13D^V5 line; (2) identifying VPS13D binding partners on Golgi membranes through proteomics screens based on proximity labeling and co-immunoprecipitation; and (3) investigating the effect of VPS13D loss on purified Golgi using lipidomics and translating findings from cell lines to conditional knockout mice and derived cells, focusing on TGN maintenance and raft-like domain formation.

Deliverables

Precise localization of VPS13D within the Golgi complex
Identification of VPS13D binding partners on Golgi membranes
Understanding of VPS13D's role in TGN maintenance and raft-like domain formation
Lipidomics data from purified Golgi
Findings from conditional knockout mice and derived cells

Technical requirements

Nocodazole ministacks
Superresolution microscopy
Immune-electron microscopy
Proteomics screens based on proximity labeling
Co-immunoprecipitation
Mass spectrometry
Lipidomics
Endogenously tagged VPS13D^V5 line
Conditional knockout mice and derived cells