Abstract
Here, we present a new approach for protein ligand screening based on the use of limited exoproteolysis coupled to MALDI-TOF mass spectrometry, combined with computational modelling and prediction of binding energies. As a test for this combined approach, we have screened a combinatorial library containing 8000 peptides (organized in 60 peptide samples) based on positional scanning format. This library is attached to a poly-Pro framework, and screened against the Abl-SH3 domain. The results obtained demonstrated the validity of the experimental and theoretical approaches in identifying better ligands and in rationalizing the changes in affinity. Exoproteolysis coupled to MALDI-TOF mass spectrometry could be used to screen complex libraries in a fast and efficient way. © 2003 Elsevier Ltd. All rights reserved.
Original language | English |
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Pages (from-to) | 1039-1048 |
Journal | Journal of Molecular Biology |
Volume | 330 |
Issue number | 5 |
DOIs | |
Publication status | Published - 25 Jul 2003 |
Keywords
- Binding constant
- Computer design
- Ligand screening
- Mass spectrometry
- SH3 domain