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

Vincent G H  Eijsink; Gerrit VRIEND; Bertus van den Burg; J Rob  van der Zee; Gerard 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

Vincent G H Eijsink, Gerrit VRIEND, Bertus van den Burg, J Rob van der Zee, Gerard Venema

Molecular Genetics

Research output: Contribution to journal › Article › Academic › peer-review

44 Citations (Scopus)

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.

Original language	English
Pages (from-to)	165-170
Number of pages	6
Journal	Protein Engineering
Volume	5
Issue number	2
DOIs	https://doi.org/10.1093/protein/5.2.165
Publication status	Published - Mar-1992

Keywords

ALPHA-HELIX
BACILLUS
HELIX DIPOLE
NEUTRAL PROTEASE
THERMOSTABILITY
BACILLUS-SUBTILIS
DIPOLE MODEL
PROTEINS
THERMOLYSIN
STABILITY
STABILIZATION
RESOLUTION
EXPRESSION
MUTATIONS
CLONING

Access to Document

10.1093/protein/5.2.165

Cite this

@article{b090ca1950584f138e4f19e0ba0dfcc5,

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

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.",

keywords = "ALPHA-HELIX, BACILLUS, HELIX DIPOLE, NEUTRAL PROTEASE, THERMOSTABILITY, BACILLUS-SUBTILIS, DIPOLE MODEL, PROTEINS, THERMOLYSIN, STABILITY, STABILIZATION, RESOLUTION, EXPRESSION, MUTATIONS, CLONING",

author = "Eijsink, {Vincent G H} and Gerrit VRIEND and {van den Burg}, Bertus and {van der Zee}, {J Rob} and Gerard Venema",

year = "1992",

month = mar,

doi = "10.1093/protein/5.2.165",

language = "English",

volume = "5",

pages = "165--170",

journal = "Protein Engineering",

issn = "0269-2139",

number = "2",

}

INCREASING THE THERMOSTABILITY OF A NEUTRAL PROTEASE BY REPLACING POSITIVELY CHARGED AMINO-ACIDS IN THE N-TERMINAL TURN OF ALPHA-HELICES. / Eijsink, Vincent G H ; VRIEND, Gerrit; van den Burg, Bertus; van der Zee, J Rob ; Venema, Gerard.

In: Protein Engineering, Vol. 5, No. 2, 03.1992, p. 165-170.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - INCREASING THE THERMOSTABILITY OF A NEUTRAL PROTEASE BY REPLACING POSITIVELY CHARGED AMINO-ACIDS IN THE N-TERMINAL TURN OF ALPHA-HELICES

AU - Eijsink, Vincent G H

AU - VRIEND, Gerrit

AU - van den Burg, Bertus

AU - van der Zee, J Rob

AU - Venema, Gerard

PY - 1992/3

Y1 - 1992/3

N2 - 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.

AB - 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.

KW - ALPHA-HELIX

KW - BACILLUS

KW - HELIX DIPOLE

KW - NEUTRAL PROTEASE

KW - THERMOSTABILITY

KW - BACILLUS-SUBTILIS

KW - DIPOLE MODEL

KW - PROTEINS

KW - THERMOLYSIN

KW - STABILITY

KW - STABILIZATION

KW - RESOLUTION

KW - EXPRESSION

KW - MUTATIONS

KW - CLONING

U2 - 10.1093/protein/5.2.165

DO - 10.1093/protein/5.2.165

M3 - Article

VL - 5

SP - 165

EP - 170

JO - Protein Engineering

JF - Protein Engineering

SN - 0269-2139

IS - 2

ER -