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Substitutions of surface amino acid residues of cutinase probed by aqueous two-phase partitioning

Berggren, Kristina; Egmond, Maarten R and Tjerneld, Folke LU (2000) In BBA - Protein Structure and Molecular Enzymology 1481(2). p.317-327
Abstract
The surface properties of a protein are often crucial for recognition and interaction with other molecules. Important functional residues can be identified by mutational analysis. There is a need for rapid methods to study protein surfaces and surface changes due to mutations. Partitioning in aqueous two-phase systems has the potential to be used in this respect since protein partitioning depends on the surface properties of the protein. The influence of surface-exposed amino acid residues in protein partitioning has been studied with cutinase variants, which differed in one or several amino acid residues as a result of site-directed mutagenesis. The solvent accessibility of the mutated residues was determined with a computer program,... (More)
The surface properties of a protein are often crucial for recognition and interaction with other molecules. Important functional residues can be identified by mutational analysis. There is a need for rapid methods to study protein surfaces and surface changes due to mutations. Partitioning in aqueous two-phase systems has the potential to be used in this respect since protein partitioning depends on the surface properties of the protein. The influence of surface-exposed amino acid residues in protein partitioning has been studied with cutinase variants, which differed in one or several amino acid residues as a result of site-directed mutagenesis. The solvent accessibility of the mutated residues was determined with a computer program, Graphical Representation and Analysis of Surface Properties. The aqueous two-phase system was composed of dextran and a random copolymer of ethylene oxide and propylene oxide. It was shown, for the first time, to what extent surface-exposed amino acid residues influence the partition coefficient in an aqueous two-phase system. The effect on partitioning could be described only taking into account solvent accessibility and type of residue substitution. The results demonstrate that the system can be used to detect conformational changes in mutant proteins since the expected effect on partitioning due to a mutation can be calculated. The aqueous two-phase system used here does indeed provide a rapid and convenient method to study protein surfaces and slight surface changes due to mutations. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Protein surface, Site-directed mutagenesis, Aqueous two-phase system, Partitioning
in
BBA - Protein Structure and Molecular Enzymology
volume
1481
issue
2
pages
317 - 327
publisher
Elsevier
external identifiers
  • scopus:0034730456
ISSN
0167-4838
DOI
10.1016/S0167-4838(00)00175-8
language
English
LU publication?
yes
id
a3fcf7c6-e7c5-4911-870b-d8c8393c99f8 (old id 125081)
date added to LUP
2007-07-04 11:43:29
date last changed
2017-01-01 07:21:21
@article{a3fcf7c6-e7c5-4911-870b-d8c8393c99f8,
  abstract     = {The surface properties of a protein are often crucial for recognition and interaction with other molecules. Important functional residues can be identified by mutational analysis. There is a need for rapid methods to study protein surfaces and surface changes due to mutations. Partitioning in aqueous two-phase systems has the potential to be used in this respect since protein partitioning depends on the surface properties of the protein. The influence of surface-exposed amino acid residues in protein partitioning has been studied with cutinase variants, which differed in one or several amino acid residues as a result of site-directed mutagenesis. The solvent accessibility of the mutated residues was determined with a computer program, Graphical Representation and Analysis of Surface Properties. The aqueous two-phase system was composed of dextran and a random copolymer of ethylene oxide and propylene oxide. It was shown, for the first time, to what extent surface-exposed amino acid residues influence the partition coefficient in an aqueous two-phase system. The effect on partitioning could be described only taking into account solvent accessibility and type of residue substitution. The results demonstrate that the system can be used to detect conformational changes in mutant proteins since the expected effect on partitioning due to a mutation can be calculated. The aqueous two-phase system used here does indeed provide a rapid and convenient method to study protein surfaces and slight surface changes due to mutations.},
  author       = {Berggren, Kristina and Egmond, Maarten R and Tjerneld, Folke},
  issn         = {0167-4838},
  keyword      = {Protein surface,Site-directed mutagenesis,Aqueous two-phase system,Partitioning},
  language     = {eng},
  number       = {2},
  pages        = {317--327},
  publisher    = {Elsevier},
  series       = {BBA - Protein Structure and Molecular Enzymology},
  title        = {Substitutions of surface amino acid residues of cutinase probed by aqueous two-phase partitioning},
  url          = {http://dx.doi.org/10.1016/S0167-4838(00)00175-8},
  volume       = {1481},
  year         = {2000},
}