An indirect method to measure trimerization constants using surface plasmon resonance
| Autor: | C. Bendtsen, Gaetano Barbato, M. Mattu |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Leucine Zippers
Zipper Biophysics Cell Biology Surface Plasmon Resonance Kinetic energy Biochemistry Fusion protein HIV Envelope Protein gp41 Recombinant Proteins Dissociation (chemistry) chemistry.chemical_compound Crystallography Monomer chemistry HIV-1 Protein Multimerization Surface plasmon resonance Isoleucine Molecular Biology |
| Zdroj: | Analytical Biochemistry. 393:126-128 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2009.06.003 |
| Popis: | The ability of surface plasmon resonance to precisely measure kinetic binding constants was exploited here to indirectly evaluate the thermodynamic dissociation trimerization constant (K(d)) of a designed chimeric protein, IZN-23, derived from an isoleucine zipper and a portion of the N-terminal helix residues of HIV-1 gp41. The method uses two monoclonal antibodies (mAbs) that display different off-rates when binding the monomeric or trimeric IZN-23. A detailed description of the data analysis strategy employed to unravel the K(d) trimerization constant from the observed off-rate kinetic values is presented, and the potential exploitation of this technique in different fields is highlighted. |
| Databáze: | OpenAIRE |
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