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Content archived on 2024-04-30

Application of novel heparan sulphate sequencing technology to the determination of binding domains for chemokines and growth factors

Objective



This proposal describes the application of novel sequencing technology for heparan sulphate and its application in the field of immunology. A demonstration project is proposed following the results of a research feasibility study and the funding of an exploratory award.
The heparan sulphates HS molecules are a class of pericellular glycosaminoglycans with fundamental roles in the control of cell growth and adhesion. Glycosaminoglycan consist of polysaccharide chains of which there are a number of different types and which are attached to the peptide chain of the molecule. In HS polysaccharide chain has a unique molecular design which takes the form of spatially-discrete, hypervariable sulphated domains (S-domains) which act as recognition motifs and also sites for sequestration and/or activation of growth factors and chemokines. Elucidation of sequence activity relationships of HS is essential for the development of therapeutic compounds for modulating the HS-mediated localization of chemokines such as IL-8, RANTES and MCP- 1 which act as migration factors for circulating lymphoid cells and inflammatory cells of the monocytoid/granulocyte lineages. The HS produced by vascular cells are the major physiological mediators of chemokine binding and the structures of these polysaccharides and their recognition specificities will be a major focus of the project. One member of the present consortium has recently shown that the binding region on HS for the chemotactic chemokine platelet factor-4 spans two S-domains with specific sequences and the spacing between these of 25 iduronic acid residues is very important. If, as seems likely, there is evidence for selectivity in HSchemokine binding the interactions may be exploited as novel targets for drugs designed to treat inflammatory and allergic diseases.
The current methods of sequencing are extremely difficult to perform and take considerable time and effort. In fact despite a lot of effort only two sites have been sequenced to date, that for anti-thrombotic and that for basic fibroblast growth factor and it is probable impossible to determine some sequences with current technology.
The application of the HS-sequencing technology in such an important area will enable the refinement of the technology in a user environment and fully demonstrate its capabilities which can then commercialized for other applications.
The main objectives of the project are therefore;
1. Establishment of technology-in users laboratories by provision of a prototype kit.
2. Identification and isolation of HS fragments with immunological functions 3. Sequence analysis of such fragments using the kits
4. Application of knowledge of sequence to immunological problems 5. Demonstration of a viable GAG sequencing technology for routine use and commercialisation of working kits.
The consortium of partners assembled for this proposal consisting of an SME, a major pharmaceutical company and leading academic laboratories is in an excellent position to accomplish all of these objectives.

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Coordinator

Wittmann Institute of Technology and Analysis of Biomolecules GmbH
EU contribution
No data
Address
21,Warthestrasse
14513 Teltrow
Germany

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Total cost

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Participants (5)

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