Advanced

Redesigning the Folding Pathway of a Model Three-helix Bundle Protein by Site-directed Mutagenesis.

Lopes, Dahabada H.J.; Chapeaurouge, Alex; Manderson, Gavin LU ; Johansson, Jonas S. and Ferreira, Sergio T. (2004) In Journal of Biological Chemistry 279(12). p.10991-10996
Abstract
Because of their limited size and complexity, de novo designed proteins are particularly useful for the detailed investigation of folding thermodynamics and mechanisms. Here, we describe how subtle changes in the hydrophobic core of a model three-helix bundle protein (GM-0) alter its folding energetics. To explore the folding tolerance of GM-0 toward amino acid sequence variability, two mutant proteins (GM-1 and GM-2) were generated. In the mutants, cavities were created in the hydrophobic core of the protein by either singly (GM-1; L35A variant) or doubly (GM-2; L35A/I39A variant) replacing large hydrophobic side chains by smaller Ala residues. The folding of GM-0 is characterized by two partially folded intermediate states exhibiting... (More)
Because of their limited size and complexity, de novo designed proteins are particularly useful for the detailed investigation of folding thermodynamics and mechanisms. Here, we describe how subtle changes in the hydrophobic core of a model three-helix bundle protein (GM-0) alter its folding energetics. To explore the folding tolerance of GM-0 toward amino acid sequence variability, two mutant proteins (GM-1 and GM-2) were generated. In the mutants, cavities were created in the hydrophobic core of the protein by either singly (GM-1; L35A variant) or doubly (GM-2; L35A/I39A variant) replacing large hydrophobic side chains by smaller Ala residues. The folding of GM-0 is characterized by two partially folded intermediate states exhibiting characteristics of molten globules, as evidenced by pressure-unfolding and pressure-assisted cold denaturation experiments. In contrast, the folding energetics of both mutants, GM-1 and GM-2, exhibit only one folding intermediate. Our results support the view that simple but biologically important folding motifs such as the three-helix bundle can reveal complex folding plasticity, and they point to the role of hydrophobic packing as a determinant of the overall stability and folding thermodynamic of the helix bundle. (Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
279
issue
12
pages
10991 - 10996
publisher
ASBMB
external identifiers
  • scopus:1642483662
ISSN
1083-351X
DOI
10.1074/jbc.M308174200
language
English
LU publication?
no
id
18dbe13f-60b6-41da-a918-348d2bb24bc9 (old id 1130525)
date added to LUP
2008-06-19 14:04:14
date last changed
2017-01-01 04:48:14
@article{18dbe13f-60b6-41da-a918-348d2bb24bc9,
  abstract     = {Because of their limited size and complexity, de novo designed proteins are particularly useful for the detailed investigation of folding thermodynamics and mechanisms. Here, we describe how subtle changes in the hydrophobic core of a model three-helix bundle protein (GM-0) alter its folding energetics. To explore the folding tolerance of GM-0 toward amino acid sequence variability, two mutant proteins (GM-1 and GM-2) were generated. In the mutants, cavities were created in the hydrophobic core of the protein by either singly (GM-1; L35A variant) or doubly (GM-2; L35A/I39A variant) replacing large hydrophobic side chains by smaller Ala residues. The folding of GM-0 is characterized by two partially folded intermediate states exhibiting characteristics of molten globules, as evidenced by pressure-unfolding and pressure-assisted cold denaturation experiments. In contrast, the folding energetics of both mutants, GM-1 and GM-2, exhibit only one folding intermediate. Our results support the view that simple but biologically important folding motifs such as the three-helix bundle can reveal complex folding plasticity, and they point to the role of hydrophobic packing as a determinant of the overall stability and folding thermodynamic of the helix bundle.},
  author       = {Lopes, Dahabada H.J. and Chapeaurouge, Alex and Manderson, Gavin and Johansson, Jonas S. and Ferreira, Sergio T.},
  issn         = {1083-351X},
  language     = {eng},
  number       = {12},
  pages        = {10991--10996},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Redesigning the Folding Pathway of a Model Three-helix Bundle Protein by Site-directed Mutagenesis.},
  url          = {http://dx.doi.org/10.1074/jbc.M308174200},
  volume       = {279},
  year         = {2004},
}