Anders Bergkvist - CV

Curriculum Vitae of Anders Bergkvist

Born 23rd of November 1969 in Skara, Sweden
Swedish citizen
Married, no children

Education
January 1996: Master of Engineering Physics at Chalmers University of Technology, Göteborg, Sweden
expected in September 2000: Ph.D. on "Structural Studies on Plastocyanin and Transhydrogenase by NMR"

Extra curricular activities at Chalmers
1992-93: President of Chalmers Aerospace Club
1993: Participation in BEST summer course in Grenoble, France
1994: COMETT-scholarship for research practice in Lisbon, Portugal

Research activities
Plastocyanin (one manuscript submitted, two more to come)
Structure, dynamics and interaction with cytochrome f

Transhydrogenase (two accepted papers, two more to come)
Assignments and global fold of domain III, domain and substrate interactions

Azurin (one or two papers to come)
Structure

Universal stress protein A, uspA
Assignments, phosphorylation site, structure, function, ...

References
Prof. Martin Billeter email: martin.billeter@bcbp.gu.se tfn: +46 31 7733925
address: Dept. of Biochemistry and Biophysics, Göteborg University

Docent Göran Karlsson email: goran.karlsson@bcbp.chalmers.se tfn: +46 31 7733917
address: Dept. of Molecular Biotechnology, Chalmers University of Technology

Prof. Thomas Nyström email: thomas.nystrom@gmm.gu.se tfn: +46 31 7732582
address: Dept. of Cell and Molecular Biology - Microbiology, Göteborg University

Lundberglab, P.O. Box 462, S-405 30 Göteborg, Sweden (applies to all)

Plastocyanin
This was my original project, started in 1995. The main issue is to determine a high-resolution structure of spinach plastocyanin. Due to several different problems, this project has been somewhat delayed.

There are already several X-ray and NMR structures of plastocyanin from other species; specifically, there are very nice NMR structures of parsley and french bean. We therefore realized, a year or so into the project, that a structure of spinach plastocyanin would not bring substantially new information to the field and that we should expand our project to include NMR relaxation analyses as well, before publication.
Later, our structure was also used in interaction studies between pc and cytf and that will also be a part of my thesis work.

Publications in prepation:
"High-resolution Solution Structure and 15N-relaxation Analysis of Spinach Plastocyanin", A Bergkvist, S Young and B G Karlsson. Will be submitted to Biochemistry during spring of 2000.

"Side-chain Interactions in the Plastocyanin-Cytochrome f Complex as Determined by 1H-NMR Chemical-shift Analysis", M Ejdebäck, A Bergkvist, G W Canters, B G Karlsson and M Ubbink. Will be submitted to Biochemistry this or next week.

"Side-chain Interactions in the E43Q/D44N and E59K/E60Q Plastocyanin double-mutants to Cytochrome f Complex as Determined by 1H-NMR Chemical-shift Analysis", A Bergkvist, M Ejdebäck, B G Karlsson and M Ubbink. Will be submitted during spring of 2000.

Related publication:
"The structure of the complex of plastocyanin and cytochrome f, determined by paramagnetic NMR and restrained rigid-body molecular dynamics", M Ubbink, M Ejdebäck, B G Karlsson and D S Bendall, Structure 6 (1998), 323-335

Transhydrogenase
This project was started in 1997 in cooperation with Prof. Jan Rydström and coworkers. Transhydrogenase is a membrane-bound enzyme that couples the reversible reduction of NADP+ by NADH to a proton translocation across the cytoplasmic membrane. The protein is composed of three domains, the NAD(H)-binding domain I, the membrane-spanning domain II and the NADP(H)-binding domain III.

We have cloned and expressed both soluble domains of E. coli transhydrogenase, ecI (dimer of 2´42.8 kDa) and ecIII (monomer of 20.4 kDa). We have published an essentially complete backbone assignment of ecIII and we also have a publication on its global fold and substrate-binding site. Currently we are working on interaction studies between ecIII and ecI, and the application of residual dipolar couplings for the structure determination of ecIII.

Existing publications:
"Sequential assignment and secondary structure analysis of the NADP(H)-binding domain of Escherichia coli transhydrogenase", C Johansson, A Bergkvist, O Fjellström, J Rydström and B G Karlsson, J. Biomol. NMR 14 (1999), 295-296.

"NMR Characterization of the NADP(H)-binding Domain of Eschericia coli Transhydrogenase; Sequential Assignment and Global Fold", C Johansson, A Bergkvist, O Fjellström, J Rydström and B G Karlsson, FEBS Lett. 458 (1999), 180-184.

Publications in prepation:
"Domain Interactions in Transhydrogenase studied by NMR and Mutagenesis", A Bergkvist, C Johansson, T Johansson, JRydström and B G Karlsson. Will be submitted to Biochemistry during autumn of 1999.

"Residual Dipolar Coupling Measurements in the NADP(H)-binding domain of Escherichia coli transhydrogenase", A Bergkvist, T Papavoine, T Johansson, M Billeter and B G Karlsson. Will be submitted during spring of 2000.

Azurin
Like plastocyanin this is a protein which was available at the department when I started my graduate studies. There are also several X-ray structures available of this protein, but as of yet there is no NMR structure. This is not my main project and it will not be included in my dissertation thesis, but I have been involved with it during the course of the project and I expect to continue playing an advisory role here.

Publications in prepation:
"Sequential Assignment of Pseudomonas Aeruginosa Azurin", J Leckner, A Bergkvist and BG Karlsson. Estimated to be submitted during spring of 2000.
"High-resolution Solution Structure of Pseudomonas Aeruginosa Azurin", J Leckner, A Bergkvist and B G Karlsson. Estimated to be submitted during autumn of 2000.

Universal Stress Protein A (UspA)
This project is a rather new NMR project and it is done in cooperation with Prof. Thomas Nyström. The major NMR work that has been done so far has been done by a diploma worker under my supervision.

UspA is an E. coli protein (15.9 kDa) which is transcriptionally induced, when the cell reaches stasis, as a function of various stress factors. The protein has been cloned and overexpresses all right in rich as well as in minimal medium (>7 mg/l). We have an (more than two years) old sample grown on rich medium which show no deterioration. However, we have had some problem with stability on proteins grown in minimal medium. A possible explanation of this is a problem with some purification step. Despite this, presumably transient, problem, we have been able to detect 15N-HSQC spectra (before the protein deteriorated) which show signal dispersion indicative of a defined fold. We also know that the protein is a dimer (total weight of 32 kDa) and that it is overexpressed with one phosphate attached to each monomer, in rich medium.

In my mind, devising a purification procedure that will leave a stable protein also in minimal medium shouldn’t be too difficult. After that, a backbone assignment would require 15N- and 13C-labeling and possibly 2H-labeling. However, a backbone assignment would then enable identification of not only secondary structure elements but also of the phosphorylation site as well as a possible ATP-binding site. Further side-chain assignments and structure calculations would require special attention to the dimeric form of the protein, but would still be quite possible.

The structure of uspA would be very interesting since no sequence homologies have been found to existing structures. The structure would also be very interesting since the protein have a well-characterized physiological, but not biochemical, function.

Related publications:
"Cloning, mapping and nucleotide sequencing of a gene encoding a universal stress protein in Escherichia coli", T Nyström and F C Neidhardt, Mol. Microbiol. 6 (1992), 3187-3198

"The Universal Stress Protein, UspA, of Escherichia coli is Phosphorylated in Response to Stasis", Freestone et al., J. Mol. Biol. 274 (1997), 318-324