Award details

Structural and functional studies of human filamin

Reference	BB/F006845/1
Principal Investigator / Supervisor	Professor Iain Campbell
Co-Investigators / Co-Supervisors
Institution	University of Oxford
Department	Biochemistry
Funding type	Research
Value (£)	319,126
Status	Completed
Type	Research Grant
Start date	01/02/2008
End date	31/01/2011
Duration	36 months

Abstract

Filamin is a large multi-domain intracellular protein that is an essential component of protein assemblies that form and break up during cell migration. Filamin cross-links actin filaments and provides binding sites for various other proteins involved in signalling. Mutations in filamin cause a variety of genetic linked diseases. In a collaborative project, involving laboratories in Yale, USA, and Jyväskylä, Finland, the structure of various filamin domains and their interactions with essential ligands, including integrins, will be studied. Filamin domains are immunoglobulin-like and are well suited for study using a dissection approach. We will focus on the region of filamin containing IgFLN domains 15-24, as this is known to contain most of the binding sites for other proteins, including integrins. Various combinations of these domains will be expressed in E.coli; their structure and interaction sites will be mapped using high resolution NMR. Particular ligands that will be studied include cytoplasmic tails of beta-integrin subunits, as well as the proteins migfilin and FILIP. Special attention will be paid to obtain information about how the protein complexes are regulated by phosphorylation and protein cleavage.

Summary

Controlled migration of cells is an important process for a variety of essential functions in the body. These include healthy processes, such as development of the embryo, wound healing and the immune response, as well as unhealthy ones, such as the uncontrolled spread of cells in cancer (metastasis). Cell migration is a complex process involving many proteins that form large dynamic assemblies. The formation and breakdown of these assemblies is carefully controlled by the cell. One key protein in the assembly is called 'integrin', a membrane-spanning receptor that adheres to molecules outside cells, thus allowing traction forces to be applied; integrins can also transmit essential signals about cell environment across the cell membrane. Another key protein is 'actin' which polymerises at the leading edge of migrating cells to drive the cell forward. Actin filaments are stabilised by a large cross-linking molecule called 'filamin'. Mutations in different parts of the filamin molecule are known to cause several different genetic diseases, implying that this molecule plays several signalling as well as structural roles in the cell. The purpose of this proposal is to investigate the structure and function of filamin so that we can better understand how it works at the molecular level. This will be done by producing a wide range of molecular fragments of this large molecule by expressing parts of the gene in bacteria. The structure of the fragments produced will be studied and the way they interact with various molecules, including integrins, will be investigated using a variety of biophysical tools. The goal is to obtain information on the atomic resolution scale (0.1nm). The results, obtained by a 'dissection' approach, will be placed in the context of the whole molecule and the whole cell using electron microscopy and intact cell studies.

Committee	Closed Committee - Biomolecular Sciences (BMS)
Research Topics	Microbiology, Structural Biology
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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