Increasing the thermostability of a neutral protease by replacing positively charged amino acids in the N-terminal turn of alpha-helices

V G Eijsink; G Vriend; B van den Burg; J R van der Zee; G Venema

doi:10.1093/protein/5.2.165

Increasing the thermostability of a neutral protease by replacing positively charged amino acids in the N-terminal turn of alpha-helices

Protein Eng. 1992 Mar;5(2):165-70. doi: 10.1093/protein/5.2.165.

Authors

V G Eijsink¹, G Vriend, B van den Burg, J R van der Zee, G Venema

Affiliation

¹ Department of Genetics, University of Groningen, Haren, The Netherlands.

PMID: 1594571
DOI: 10.1093/protein/5.2.165

Abstract

The 247-260 and 289-299 alpha-helices of Bacillus subtilis neutral protease have a lysine in their N-terminal turn. These lysines were replaced by Ser or Asp in order to improve electrostatic interactions with the alpha-helix dipole. After replacing Lys by Ser at positions 249 or 290, the thermostability of the enzyme was increased by 0.3 and 1.0 degrees C, respectively. The Asp249 and Asp290 mutants exhibited a stabilization of 0.6 and 1.2 degrees C, respectively. The results show the feasibility of stabilizing enzymes by introducing favourable residues at the end of alpha-helices.

Publication types

Comparative Study

MeSH terms

Amino Acid Sequence
Bacillus subtilis / enzymology
Bacillus subtilis / genetics*
Enzyme Stability / genetics
Metalloendopeptidases / genetics*
Metalloendopeptidases / metabolism
Models, Molecular
Molecular Sequence Data
Mutagenesis, Site-Directed
Mutation
Protein Conformation
Protein Denaturation / genetics

Substances

Metalloendopeptidases
microbial metalloproteinases