INCREASING THE THERMOSTABILITY OF A NEUTRAL PROTEASE BY REPLACING POSITIVELY CHARGED AMINO-ACIDS IN THE N-TERMINAL TURN OF ALPHA-HELICES

Authors
EIJSINK VGH VRIEND G VANDENBURG B VANDERZEE JR VENEMA G
Citation
Vgh. Eijsink et al., INCREASING THE THERMOSTABILITY OF A NEUTRAL PROTEASE BY REPLACING POSITIVELY CHARGED AMINO-ACIDS IN THE N-TERMINAL TURN OF ALPHA-HELICES, Protein engineering, 5(2), 1992, pp. 165-170
Citations number
44
Journal title
Protein engineering → ACNP
ISSN journal
02692139
Volume
5
Issue
2
Year of publication
1992
Pages
165 - 170
Database
ISI
SICI code
0269-2139(1992)5:2<165:ITTOAN>2.0.ZU;2-G
Abstract
The 247-260 and 289-299 alpha-helices of Bacillus subtilis neutral pro tease have a lysine in their N-terminal turn. These lysines were repla ced 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-d egrees-C, respectively. The Asp249 and Asp290 mutants exhibited a stab ilization 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.