Allosteric Control of Ligand Binding Affinity Using Engineered Conformation-Specific Effector Proteins
| Autor: | John H Heithaus, Erica M. Duguid, Shahir S. Rizk, Anthony A. Kossiakoff, Andrew Sandstrom, Marcin Paduch |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Models
Molecular 0303 health sciences Phage display biology Protein Conformation Synthetic antigen 030302 biochemistry & molecular biology Allosteric regulation Cooperative binding Maltose binding Protein engineering Ligand (biochemistry) Ligands Protein Engineering Article 03 medical and health sciences Allosteric enzyme Biochemistry Allosteric Regulation Structural Biology Mutation biology.protein Molecular Biology 030304 developmental biology Protein Binding |
| Zdroj: | Nature structural & molecular biology |
| ISSN: | 1545-9985 1545-9993 |
| Popis: | We describe a phage display methodology for engineering synthetic antigen binders (sABs) that recognize either the apo or the ligand-bound conformation of maltose-binding protein (MBP). sABs that preferentially recognize the maltose-bound form of MBP act as positive allosteric effectors by substantially increasing the affinity for maltose. A crystal structure of a sAB bound to the closed form of MBP reveals the basis for this allosteric effect. We show that sABs that recognize the bound form of MBP can rescue the function of a binding-deficient mutant by restoring its natural affinity for maltose. Furthermore, the sABs can enhance maltose binding in vivo, as they provide a growth advantage to bacteria under low-maltose conditions. The results demonstrate that structure-specific sABs can be engineered to dynamically control ligand-binding affinities by modulating the transition between different conformations. |
| Databáze: | OpenAIRE |
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