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Evolution of a carbohydrate binding module into a protein-specific binder

Cicortas Gunnarsson, Lavinia LU ; Dexlin Mellby, Linda LU ; Nordberg Karlsson, Eva LU orcid ; Holst, Olle LU and Ohlin, Mats LU orcid (2006) In Biomolecular Engineering 23(2-3). p.111-117
Abstract
A carbohydrate binding module, CBM4-2, derived front the xylanase (Xyn 10A) of Rhodothermus marinus has been used as a scaffold for molecular diversification. Its binding specificity has been evolved to recognise a quite different target, a human monoclonal IgG4. In order to understand the basis for this drastic change in specificity we have further investigated the target recognition of the IgG4-specific CBMs. Firstly, we defined that the structure target recognised by the selected CBM-variants was the protein and not the carbohydrates attached to the glycoprotein. We also identified key residues involved in the new specificity and/or responsible for the swap in specificity, from xylan to human IgG4. Specific changes present in all these... (More)
A carbohydrate binding module, CBM4-2, derived front the xylanase (Xyn 10A) of Rhodothermus marinus has been used as a scaffold for molecular diversification. Its binding specificity has been evolved to recognise a quite different target, a human monoclonal IgG4. In order to understand the basis for this drastic change in specificity we have further investigated the target recognition of the IgG4-specific CBMs. Firstly, we defined that the structure target recognised by the selected CBM-variants was the protein and not the carbohydrates attached to the glycoprotein. We also identified key residues involved in the new specificity and/or responsible for the swap in specificity, from xylan to human IgG4. Specific changes present in all these CBMs included mutations not introduced in the design of the library from which the specific clones were selected. Reversion of such mutations led to a complete loss of binding to the target molecule, suggesting that they are critical for the recognition of human IgG4. Together with the mutations introduced at will, they had transformed the CBM scaffold into a protein binder. We have thus shown that the scaffold of CBM4-2 is able to harbour molecular recognition for either carbohydrate or protein structures. (c) 2005 Elsevier B.V. All rights reserved. (Less)
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author
; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
molecular engineering, protein scaffold, phage-display, mutant, binding specificity, combinatorial library
in
Biomolecular Engineering
volume
23
issue
2-3
pages
111 - 117
publisher
Elsevier
external identifiers
  • wos:000237050100004
  • pmid:16427804
  • scopus:33645457940
ISSN
1389-0344
DOI
10.1016/j.bioeng.2005.12.002
project
Designed carbohydrate binding modules and molecular probes
language
English
LU publication?
yes
id
8d3c1c0c-486b-4738-91d7-26d0ab6fa819 (old id 410957)
date added to LUP
2016-04-01 16:06:18
date last changed
2021-09-22 03:43:14
@article{8d3c1c0c-486b-4738-91d7-26d0ab6fa819,
  abstract     = {A carbohydrate binding module, CBM4-2, derived front the xylanase (Xyn 10A) of Rhodothermus marinus has been used as a scaffold for molecular diversification. Its binding specificity has been evolved to recognise a quite different target, a human monoclonal IgG4. In order to understand the basis for this drastic change in specificity we have further investigated the target recognition of the IgG4-specific CBMs. Firstly, we defined that the structure target recognised by the selected CBM-variants was the protein and not the carbohydrates attached to the glycoprotein. We also identified key residues involved in the new specificity and/or responsible for the swap in specificity, from xylan to human IgG4. Specific changes present in all these CBMs included mutations not introduced in the design of the library from which the specific clones were selected. Reversion of such mutations led to a complete loss of binding to the target molecule, suggesting that they are critical for the recognition of human IgG4. Together with the mutations introduced at will, they had transformed the CBM scaffold into a protein binder. We have thus shown that the scaffold of CBM4-2 is able to harbour molecular recognition for either carbohydrate or protein structures. (c) 2005 Elsevier B.V. All rights reserved.},
  author       = {Cicortas Gunnarsson, Lavinia and Dexlin Mellby, Linda and Nordberg Karlsson, Eva and Holst, Olle and Ohlin, Mats},
  issn         = {1389-0344},
  language     = {eng},
  number       = {2-3},
  pages        = {111--117},
  publisher    = {Elsevier},
  series       = {Biomolecular Engineering},
  title        = {Evolution of a carbohydrate binding module into a protein-specific binder},
  url          = {http://dx.doi.org/10.1016/j.bioeng.2005.12.002},
  doi          = {10.1016/j.bioeng.2005.12.002},
  volume       = {23},
  year         = {2006},
}