Publication

Model building of a thermolysin-like protease by mutagenesis

Frigerio, F., Margarit, N. V., Nogarotto, R., Grandi, G., Vriend, G., Hardy, F., Veltman, OR., Venema, G. & Eijsink, VGH., Mar-1997, In : Protein Engineering. 10, 3, p. 223-230 8 p.

Research output: Contribution to journalArticleAcademicpeer-review

  • F Frigerio
  • [No Value] Margarit
  • R Nogarotto
  • G Grandi
  • G Vriend
  • F Hardy
  • OR Veltman
  • G Venema
  • VGH Eijsink

The present study concerns the use of site-directed mutagenesis experiments to optimize a three-dimensional model of the neutral protease of Bacillus subtilis (NP-sub), An initial model of NP-sub was constructed using the crystal structures of the homologous neutral proteases of Bacillus thermoproteolyticus (thermolysin) and Bacillus cereus as templates, The largest portion of NP-sub could be modelled satisfactorily, using standard techniques, but several surface-located regions could only be modelled with a high degree of uncertainty, In order to make the model more reliable in these regions a 'model building by mutagenesis' approach was adopted, Mutations were designed such that their effect on thermal stability could indicate how their local environment should be modelled, This approach provided insight in the local structure of several regions in NP-sub that were hard to model on the basis of homology with the two known structures alone.

Original languageEnglish
Pages (from-to)223-230
Number of pages8
JournalProtein Engineering
Volume10
Issue number3
Publication statusPublished - Mar-1997

    Keywords

  • Bacillus subtilis, homology modelling, mutagenesis, neutral protease, surface loops, SUBTILIS NEUTRAL PROTEASE, HINGE-BENDING MOTION, BACILLUS-SUBTILIS, THERMAL-STABILITY, SEQUENCE, CEREUS, GENE, RESOLUTION, PROTEINS, CLONING

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