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

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Author

Summary, in English

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.

Publishing year

2006

Language

English

Pages

111-117

Publication/Series

Biomolecular Engineering

Volume

23

Issue

2-3

Document type

Journal article

Publisher

Elsevier

Topic

  • Industrial Biotechnology
  • Immunology in the medical area

Keywords

  • molecular engineering
  • protein scaffold
  • phage-display
  • mutant
  • binding specificity
  • combinatorial library

Status

Published

Project

  • Designed carbohydrate binding modules and molecular probes

ISBN/ISSN/Other

  • ISSN: 1389-0344