Competence of the organizing groups

Protein Crystallography, Prof. Rik Wierenga

An extensive infrastructure for protein crystallographic studies has been built up, consisting of five components: teaching/training, protein characterization, protein crystallization, data collection and crystallographic computing. The protein crystallographic research is complemented by enzymological studies and detailed structure analysis studies. The enzymological characterizations are aimed at a quantitative description of important properties of our enzymes and include enzyme kinetic studies, protein stability studies and affinity studies (Alahuhta et al., 2008; Alahuhta et al, 2008; Haapalainen, Merilainen et al. 2007; Merilainen et al, manuscript in preparation). The protein crystallographic studies concern three classes of enzymes: (i) triosephosphate isomerases, (ii) CoA-dependent enzymes using thioester chemistry and (iii) prolyl-4-hydroxylases. In each of these projects important new insight into the reaction mechanisms has been achieved. In specific cases we use our committed interest and expertise on biocatalysis for the development of new enzymes (TIM) or new transitionstate analogues (thiolase).

Websites:

Selected papers:

  • Alahuhta et al. Structure-based protein engineering efforts with an inactive monomeric TIM variant: the importance of a single point mutation for generating a competent active site. PEDS 1-10, 2008.
  • Alahuhta et al. The A178L mutation in the C-terminal hinge of the flexible loop-6 of dimeric and monomeric TIM favours the closed active site geometry. Acta Crystallogr D Biol Crystallogr. D64, 178-188, 2008.
  • Koski, et al. The active site of an algal prolyl-4-hydroxylase has a large structural plasticity. JBC, 282, 37112-37123, 2007.
  • Haapalainen, A.M., Meriläinen, G., et al. Crystallographic and kinetic studies of human mitochondrial acetoacetyl-CoA thiolase (T2): the importance of potassium and chloride ions for its structure and function. Biochemistry, 46, 4305-4321, 2007.
  • Casteleijn, M.G., Alahuhta, M., et al. Functional role of the conserved active site proline of triosephosphate isomerase. Biochemistry, 45, 15483-15494, 2006.