Evolution of a carbohydrate binding module into a protein-specific binder
Cicortas Gunnarsson, Lavinia LU ; Dexlin Mellby, Linda LU ; Nordberg Karlsson, Eva LU
; Holst, Olle
LU
and Ohlin, Mats
LU
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/410957
- author
- Cicortas Gunnarsson, Lavinia
LU
; Dexlin Mellby, Linda
LU
; Nordberg Karlsson, Eva
LU
; Holst, Olle
LU
and Ohlin, Mats
LU
- organization
- publishing date
- 2006
- 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
- 2022-01-28 17:22:38
@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}},
keywords = {{molecular engineering; protein scaffold; phage-display; mutant; binding specificity; combinatorial library}},
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}},
}