Molecular organisation of the secretory and endocytic pathways

Lead Research Organisation: University of Manchester
Department Name: Life Sciences

Abstract

All animal cells release proteins into and take up proteins from the surrounding fluid. They also transport proteins between internal compartments, each of which has a unique composition and a specific function. Transport out of and into cells, and between internal compartments, is crucial for the normal functioning of cells, as well as the inter-cellular communication that is essential in all animals. Defects in these transport steps are associated with many diseases. It is therefore important we understand how this transport operates at the molecular level. The aim of the work in this application is to apply the technique of high-resolution electron microscopy, combined with a variety of modern molecular approaches, to increase our understanding of a number of key transport steps. This will allow us to gain insights into how the transport pathways function in a healthy cell, and consequently how defects in particular aspects of this transport may contribute to disease.

Technical Summary

The secretory and endocytic pathways are of fundamental importance to eukaryotic cells. The secretory pathway is responsible for the synthesis of new lipid, secretory and membrane proteins, and their subsequent delivery to the correct final destination, either inside or outside the cell. The endocytic pathway is responsible for the internalisation of proteins and their sorting to the appropriate intracellular compartment or recycling back to the cell surface. Although much progress has been made in identifying the machinery that makes these pathways function correctly, there is much we still do not understand. The focus of the four groups named in this proposal is to develop this understanding by focussing on key aspects of both pathways. The aim of this application is to provide electron microscopy support for the named research groups, allowing them to pursue a variety of related projects that rely on high-resolution analysis of protein distribution or cellular morphology. By combining electron microscopy with a number of established molecular approaches it will be possible to address specific questions that would otherwise be intractable. This work will increase our understanding of the molecular organisation of the secretory and endocytic pathways, and how defects in specific trafficking components may contribute to disease.

Publications

10 25 50
 
Title Antibodies to Golgi apparatus proteins 
Description Polyclonal antibodies to a variety of Golgi apparatus proteins. These are ZFPL1, GM130, GMAP-210. 
Type Of Material Antibody 
Year Produced 2007 
Provided To Others? Yes  
Impact It has helped us to publish several manuscripts and contributed to the research of other labs in the UK and abroad. 
 
Title Antibodies to Golgi proteins 
Description Antibodies to Golgi proteins TMF1, ZFPL1 and GMAP210 
Type Of Material Antibody 
Year Produced 2008 
Provided To Others? Yes  
Impact Not yet 
 
Title Cell lines stably expressing Golgi apparatus proteins 
Description Cell lines stably expressing GFP-tagged versions of the Golgi proteins ZFPL, GMAP210 and GM130. 
Type Of Material Cell line 
Year Produced 2008 
Provided To Others? Yes  
Impact None yet 
 
Title HeLa derived stable cell lines 
Description HeLa derived cell lines stably expressing versions of the novel Golgi protein ZFPL1. 
Type Of Material Cell line 
Provided To Others? No  
Impact Allowed us to publish EMBO J paper on functions of ZFPL1 
 
Title New solid phase protein binding assay 
Description It is a solid-phase binding assay for the analysis of protein-protein interactions. 
Type Of Material Technology assay or reagent 
Year Produced 2008 
Provided To Others? Yes  
Impact It allowed us to perform more quantitative binding assays, contributing to publications. 
 
Title Viral secretion marker for cell biology 
Description It is a viral vector for the expression of tsO45G into a multitude of mammalian cells. TsO45G is a commonly used secretory marker. Our vector allows efficient delivery into cells that are undergoing a wide variety of treatments. 
Type Of Material Technology assay or reagent 
Year Produced 2007 
Provided To Others? Yes  
Impact It helped us to publish 2 manuscripts on Golgi trafficking. 
 
Description Article for Biological Sciences Review 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Philip Woodman has written an article for Biological Sciences Review, a lay publication aimed at 6th form school pupils.

None
Year(s) Of Engagement Activity 2009