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Spider Web Proteins for Medicine

#1033


Obtaining and Strucutal-Functional Investigations of Spider Web Recombinant Proteins and their Synthetic Analogs for the Purpose of the Development of New Materials Designed for Medical Application

Tech Area / Field

  • MED-RAD/Radiomedicine/Medicine

Status
8 Project completed

Registration date
08.07.1997

Completion date
04.12.2000

Senior Project Manager
Pobedimskaya D D

Leading Institute
State Scientific Center of Genetics and Selection of Industrial Microorganisms (GosNIIGenetica), Russia, Moscow

Supporting institutes

  • State Research Center for Applied Microbiology, Russia, Moscow reg., Obolensk\nScientific Research Center "Uglechimvolokno", Russia, Moscow reg., Mytischi

Collaborators

  • University of Michigan, USA, MI, Ann Arbor

Project summary

Up till now there has not been constructed a material simultaneously possessing properties of natural silk fibres, such as (1) elasticity (ability to preserve elastic behavior under 30-40% expansion), (2) amazing strength (the tensile strength of the strongest dragline silk - 200 000 psi - is several times greater than steel and of the same order as Kevlar), (3) stability to large variations of temperature and humidity and (4) low level of immunogenity. Due to its physico-chemical and mechano-chemical properties and special architectonics) fibrous proteins may be a basis for creation of perspective materials of medical purposes for regeneration of a skin recover and creation of artificial sutures, ligament and tendon in microsurgeries and rehabilitation surgery.

The main goal of the proposed project is creation of highly effective bacterial strains-producers of recombinant silk proteins and their synthetic analogous, study of their physical and chemical and biomechanical properties and study of the approaches to creation of medical materials on the basis of recombinant polypeptides.

The technical approach and methodology


1. The oligonucleotide chemical synthesis using automatic DNA synthesizer.
2. Amplification of DNA fragments due to polymerize chain reaction for constructing of model recombinant web protein genes followed by their molecular cloning.
3. Obtaining of high efficient expression of native web protein genes and their artificial analogs due to molecular cloning of corresponding DNA fragments in the specialized prokaryotic expression vectors.
4. Exploiting of different methods of biochemistry and protein chemistry, HPLC and other chromatography procedures, in particular, for development of analytical and large-scale recombinant polypeptides extraction and purification.
5. Physico-chemical studies of recombinant proteins on the basis of IR-spectroscopy, CD, NMR-relaxation, X-ray with rotated and and synchrotronic irradiation and neutron scattering, as well as dilatometrical and mechano-chemical investigations.
6. Theoretical analysis of structural-functional relationships in recombinant polypeptides and their native analogs provided on the basis of establishing of homologies and differences between investigated and other proteins of web silk; investigation of periodicity and finding of those amino acid residues which are responsible for establishment of periodical structure; computing of conformations of asymmetrical units corresponded to different types of periodicities, due to exploiting of molecular mechanics and graphics algorithms: filling in and analyzing of data bank of web-silk proteins' sequences, structures and properties; obtaining of theoretical recommendations for arranging of model periodical structures of recombinant proteins with determined physico-chemical properties.
7. Investigation of possibilities of polymeric films and fibrous threads formation from solutions of recombinant polypeptides. These experiments would be provided, in particular, by passing of protein solutions through the narrow opening under changing of pressure and other parameters of the process followed by analysis of films and threads strength and elasticity.
8. Medico-biological trials of materials designed from recombinant proteins and their analogs based on analysis of biocompatibility of dried protein films and threads being in contact with inquired and healthy tissues of laboratory animals.

The main expected results


- Obtained bacterial strains producing recombinant web proteins and their synthetic analogous are expected to be of the important practical and potential commercial value.
- Obtained information about physico-chemical properties both of original spider web proteins as well as modeling recombinant polypeptides will have theoretical significance for solving the fundamental problem of interconnection between structure and function for fibrous proteins.
- The development of the adequate approach for the modeling of fibrous proteins with necessary physical and chemical properties will have theoretical and practical importance.
- Laboratory testing on the animals of the samples of medical preparations in film form for the wound treatment developed on the basis of new materials.
- The possibility of the usage of the threads constructed from recombinant proteins as new generation materials for the surgery usage will be evaluated.
- The study of potential for design of new drugs and biocompatible materials on the basis of fibrous proteins will be performed.


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