Position: "Forskare" (LU)
Project: Protein structure and dynamics
Funding ends: 31 December, 2000
Two main projects are being pursued: [1] High-throughput protein folding and [2] Structure, function and dynamics of a synaptic efficacy modulator.
[1] The first project deals with investigations of the mechanism by which the protein sequence information in one dimension is converted into the three-dimensional structure of a protein, i.e. the protein folding problem. The goal of the project is to optimise the conditions of protein refolding by varying a large range of conditions in an organised and methodical way, a so-called "high-throughput" approach. The applications of this project are foreseen to be useful in improving production of proteins from the laboratory scale up to industrial scale. This project builds on studies carried out previously on the stability and folding mechanism of a domain of calmodulin, TR2C. That study involved measurement of hydrogen exchange rates of the protein under various conditions in order to analyse the order of folding events. Those studies were completed during 2000 and the results are being prepared for publication. Initial investigations of the high-throughput project are promising but the project suffers from a lack of personnel which hinders its successful completion.
[2] The second project involves the determination of the structure of a 190 amino acid protein identified as a regulator of neural efficacy in Drosophila and later also isolated from mammals (rat, human). The protein has partial homology with the recently characterised photoreceptor proteins, recoverin and neurocalcin. In addition to sequence similarity, they shares properties of N-terminal acylation and calcium-dependent activity. The protein has been produced in large quantities with both 15N and 13C labeling by my collaborators, Prof. Olaf Pongs and Dr. Alexander Hauenschild of Hamburg University. The complete resonance assignment was carried out using data from a battery of 3- and 4-dimensional triple resonance experiments on our 600 MHz Varian Unity+ NMR spectrometer. An article on these results has been published in early 2000 (1). Structure calculation is also underway and we hope to complete the high-resolution structure of the protein in the near future. As with project 1, these studies also suffer from a lack of manpower and will be difficult to complete.
Summary
Both projects carried out during the term of the SBNet
grant have shown great promise and have been given
support from the research councils in the form of grants
for materials. At the time of initiation, my research
group was composed of 2.5 persons (including PI, one
post-doc and a shared graduate student.). Since then,
the group has been decimated and today only the PI
remains to drive two projects, in addition to his other
teaching, administrative and computer system-administration
responsibilities. This situation can be partially attributed
to recent changes in the leadership structure of this
department and a reduced level of enthusiasm for structural
biology, in particular for NMR as a tool in structural
biology, in the new leadership. Running research projects
can be compared to gardening in that it requires sunlight
(in the form of ideas), water and nourishment (in the
form of funding), and gardeners (in the form of students
and post-docs). Sadly, the last has been withheld and
the garden has suffered. I have no choice but to seek
new fields to tend.
Funding (2000)
Project 1: TFR (high-throughput protein folding)300.000
Project 2: SSF special program in cellular signalling300.000
Publications
(1) Kragelund, B. B., Hauenschild, A., Carlström,
G., Pongs, O. and Finn, B. E. (2000) 1H, 13C and 15N
resonance assignments, secondary structure and fold
of Ca2+-frequenin, a synaptic efficacy modulator. J.
Biomol. NMR. 16, 85-86.
(2) Drakenberg, T., Finn, B. E. and Forsén, S.
(2001) Calcium. In Biological Inorganic Chemistry:
Structure and Reactivity. (Valentine, J., Bertini,
I. and Gray, H.B., eds.) Marcel Dekker, New York, in
press.
(3) Muranyi, A. and Finn B. E. (2001) Calcium and its
enzymes. In Handbook on Metalloproteins, 37, (Bertini,
I., Sigel, A. and Sigel, H., eds.) Marcel Dekker, New
York, in press.
Conferences
* Bioinformatics 2000, April 27-29, 2000, Helsingör,
Denmark
* SBNet-sponsored ARIA workshop, Göteborg
Latest update at 12 March, 2001.