| title: | Development of techniques for large-scale biosynthesis of 13C/2H/15N labeled peptides and proteins. Studies of structural and dynamical aspects of peptide hormone action by NMR in solution and in condensed phase. |
|---|---|
| reg no: | ETF5138 |
| project type: | Estonian Science Foundation research grant |
| subject: |
1.5. Biophysics 1.8. Molecular Biology 2.7. Biotechnology, Food and Drink Technology |
| status: | completed |
| institution: | Institute of Chemical Physics and Biophysics |
| head of project: | Kalju Vanatalu |
| duration: | 01.01.2002 - 31.12.2005 |
| description: | The goal of the project is elucidation of structural and dynamical aspects of peptide hormone (motilin and galanin) action and their interactions with partner molecules by sophisticated NMR techniques in solution and in condensed phase. The absolute prerequisite for mentioned studies is elaboration of general approach for effective biosynthesis of isotopically (2H/13C/15N) labelled peptides and proteins in preparative amounts (10 mg). As isotopical labelling of peptides sets specific requirements for the cultivation procedure, specific optimization procedures will be elaborated for optimization of the yield in deuterated growth medium. Next step of present project is connected with purification, analysis and crystallization of expressed products where specific methods for purification of recombinant peptides and crystallization of peptides and small proteins will be applied. Highly purified, isotopically labelled peptides will be studied by NMR in solution and in solid state. Peptide crystals, if suitable, will be studied also by X-ray diffraction techniques. Results awaited from biotechnological and biochemical steps of present project are of great value in form of methods for expression of the isotopically labeled proteins, their purification and crystallization. Biophysical measurements will yield a fundamental information about structural and dynamical aspects of peptide behaviour in solution and in condensed phase whereas solid state NMR experiments use several novel approaches at a top technological level. These studies in complex will enable to learn more about the basic processes of the transmission of signals across membranes which is of great fundamental and strategic value in medicine and pharmacological industry. Project is both prophoundly interdisciplinary, taking advantage of intermediate feedback between object labeling/preparation and NMR resolution optimization, and deeply coordinated between different national and international groups, leading to synergestic effects. |
| project group | ||||
|---|---|---|---|---|
| no | name | institution | position | |
| 1. | Jüri Jarvet | NICPB/Stockholm University | sen. researcher | |
| 2. | Jekaterina Kazantseva | Tallinn Technical University | doct. student | |
| 3. | Peep Palumaa | Tallinn Technical University | sen. reseacher | |
| 4. | Ago Samoson | NICPB | director | |
| 5. | Katrin Tomson | NICPB | doct. student | |
| 6. | Kalju Vanatalu | NICPB | Senior Researcher | |