Title:The targeting and biosynthesis of membrane and secretory proteins.
Our objective is to provide a detailed understanding of the biosynthesis of membrane and secretory proteins. The focus will be: 1) components 2) molecular mechanisms and 3) regulation. A detailed molecular understanding of these key issues is currently lacking. Furthermore, although membrane protein biosynthesis is directly relevant to exploiting the cell factory, little effort has been made to establish whether this process can be engineered. We will achieve our objective by focusing the research efforts of individual teams into a co-ordinated research programme. The teams provide several different biological systems, enabling information and technology transfer, and represent all of the disciplines crucial to the project: biochemistry; cell biology; microbiology; structural biology; biocomputing and commercial exploitation.
Description of the work: (please write approx. 20 lines)
A detailed understanding of the structure, function and regulation of SRP is central to understanding membrane protein biosynthesis. Subunits of SRP and the SRP receptor from different organisms will be over-expressed using several strategies and purified. The components will be reconstituted into complexes and tested for function. Functional particles will be crystallised and their high resolution structures solved. The structures will be analysed with and without specific regulatory components allowing the role of these components to be established. The molecular basis for membrane protein integration will be established by reconstituting the process in vitro and identifying components that associate with the proteins during membrane insertion. A systematic analysis will be used to dissect out "stage specific" interactions occurring at discreet steps during membrane integration. The endoplasmic reticulum (ER) mediates the N-glycosylation of newly synthesised proteins and the retention of misfolded proteins prior to their ultimate degradation. Both processes have important consequences for the production of biologically active membrane proteins and their subsequent appearance at the cell surface. Established in vitro approaches will be used to identify the components responsible for regulating key check points during the N-glycosylation and ER retention of membrane proteins, together with the molecular basis for their function. We will endeavour to exploit our detailed knowledge of membrane protein biosynthesis in a commercial framework. This will be achieved by engineering key components and establishing the effect that this has upon the production of commercially relevant, biologically active, target proteins. The whole project will be underpinned by the use of biocomputing and data base exploitation to both identify and characterise membrane targeting and integration components and predict their structure and function.
Structural determination of SRP and SRP sub-complexes
Identifying the molecular basis of SRP function and the regulation of SRP
Elucidating molecular mechanisms of membrane protein integration
Establishing the molecular basis for the regulation of N-glycosylation
Identifying components responsible for the quality control of misfolded membrane proteins
Establishing whether specific components can be engineered to enhance membrane and secretory protein production and downstream exploitation
School of Biological Sciences
University of Manchester
M13 9PT, Manchester, UK
EMBL Grenoble Outstation
c/o ILL, 156X
38042 Grenoble Cedex 9, France
Dept. Molecular Microbiology
Vrije Universiteit, De Boelelaan 1087
1081HV Amsterdam, Netherlands
Dept. of Molecular Structural Biology
BZH, University of Heidelberg
INF 328, 69120 Heidelberg. Germany.
Tel: 0049-6221-387 274
Fax: 0049-6221-387 306
Umeå Center for Molecular Pathogenesis,
Umeå University, S-90187 Umeå. Sweden
Tel: 0046 90-7856782)
Fax: 0046 90-778007)
Département de biologie cellulaire
Université de Genève
1211 Geneva 4, Switzerland
Tel: 0041-22-702 6724,
Fax: 0041-22-702 6442,
Im Neuenheimer Feld 282
Institute Mol. Biol. & Biotech.
711 10 Iraklio-Crete, GREECE.
Fax: (00)30-81- 391-166
AstraZeneca Structural Chemistry Laboratory
S-431 83, Mölndal, Sweden
Tel: (00)46-31-77 6 18 30
Fax: (00)46-31-77 6 38 35
Department of Medical Biochemistry
University of Goteborg, Medicinareg. 9A,
Box 440, S-405 30 SWEDEN.
Tel: 0046 31 773 34 68
Fax: 0046 31 41 61 08