Download MendeleyCation binding to a Bacillus (1,3-1,4)-beta-glucanase. Geometry, affinity and effect on protein stability
Keitel, T., Meldgaard, M., Heinemann, U.(1994) Eur J Biochem 222: 203-214
- PubMed: 8200344
- DOI: https://doi.org/10.1111/j.1432-1033.1994.tb18858.x
- Primary Citation of Related Structures:
1GLH - PubMed Abstract:
The hybrid Bacillus (1,3-1,4)-beta-glucanase H(A16-M), consisting of 16 N-terminal amino acids derived from the mature form of the B. amyloliquefaciens enzyme and of 198 C-proximal amino acids from the B. macerans enzyme, binds a calcium ion at a site at its molecular surface remote from the active center [T. Keitel, O. Simon, R. Borriss & U. Heinemann (1993) Proc. Natl Acad. Sci. USA 90, 5287-5291]. X-ray diffraction analysis at 0.22-nm resolution of crystals grown in the absence of calcium and in the presence of EDTA shows this site to be occupied by a sodium ion. Whereas the calcium ion has six oxygen atoms in its coordination sphere, two of which are from water molecules, sodium is fivefold coordinated with a fifth ligand belonging to a symmetry-related protein molecule in the crystal lattice. The affinity of H(A16-M) for calcium over sodium has been determined calorimetrically. Calcium binding stabilizes the native three-dimensional structure of the protein as shown by guanidinium chloride unfolding and thermal inactivation experiments. The enhanced enzymic activity of Bacillus beta-glucanases at elevated temperatures in the presence of calcium ions is attributed to a general stabilizing effect by the cation.
Organizational Affiliation:
Institut für Kristallographie, Freie Universität Berlin, Germany.
