title: Stabilization of Bacillus stearothermophilus neutral protease by introduction of prolines.
abstract: The thermostability of neutral proteases has been shown to depend on autolysis which presumably occurs in flexible regions of the protein. In an attempt to rigidify such a region in the neutral protease of Bacillus stearothermophilus, residues in the solvent-exposed 63-69 loop were replaced by proline. The mutations caused large positive (Ser-65-->Pro, Ala-69-->Pro) or negative (Thr-63-->Pro, Tyr-66-->Pro) changes in thermostability, which were explained on the basis of molecular modelling of the mutant proteins. The data show that the introduction of prolines at carefully selected positions in the protein can be a powerful method for stabilization.
authors.affiliation: Department of Genetics, University of Groningen, Haren, The Netherlands.
authors.forename: F
authors.initials: F
authors.lastname: Hardy
authors.forename: G
authors.initials: G
authors.lastname: Vriend
authors.forename: O R
authors.initials: OR
authors.lastname: Veltman
authors.forename: B
authors.initials: B
authors.lastname: van der Vinne
authors.forename: G
authors.initials: G
authors.lastname: Venema
authors.forename: V G
authors.initials: VG
authors.lastname: Eijsink
mesh_terms: D001426:Bacterial Proteins; D010450:Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D004795:Enzyme Stability; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D006358:Hot Temperature; D009154:Mutation; D011392:Proline / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation
chemical_list: D001426:Bacterial Proteins; D011392:Proline; D010450:Endopeptidases; C065625:neutral protease, Bacillus stearothermophilus