Improved molecular recognition of Carbonic Anhydrase IX by polypeptide conjugation to acetazolamide
| Autor: | Zoë Fisher, Jie Yang, Lars Baltzer, Wolfgang Knecht, Katarina Koruza |
|---|---|
| Přispěvatelé: | Department of Bio-engineering Sciences |
| Rok vydání: | 2017 |
| Předmět: |
Stereochemistry
Clinical Biochemistry Pharmaceutical Science 010402 general chemistry Crystallography X-Ray 01 natural sciences Biochemistry Small Molecule Libraries Molecular recognition Drug Discovery medicine Humans Amino Acid Sequence Surface plasmon resonance Carbonic Anhydrase IX Molecular Biology Molecular Structure 010405 organic chemistry Chemistry Organic Chemistry Surface Plasmon Resonance 0104 chemical sciences Acetazolamide Cell and molecular biology Molecular Medicine Peptides medicine.drug Protein Binding |
| Zdroj: | Bioorganicmedicinal chemistry. 25(20) |
| ISSN: | 1464-3391 |
| Popis: | The small molecule inhibitor acetazolamide (AZM) was conjugated to a set of designed polypeptides and the resulting conjugates were evaluated for their affinity to Human Carbonic Anhydrase II (HCA II) using surface plasmon resonance. The dissociation constant of the AZM-HCA II complex was 38nM and that of the AZM conjugated polypeptide (4-C10L17-AZM) to HCA II was found to be 4nM, an affinity enhancement of a factor of 10 due to polypeptide conjugation. For Human Carbonic Anhydrase IX (HCA IX) the dissociation constant of AZM was 3nM, whereas that of the 4-C10L17-AZM conjugate was 90pM, a 33-fold affinity enhancement. This dramatic affinity increase due to polypeptide conjugation was achieved for a small molecule ligand with an already high affinity to the target protein. This supports the concept that enhancements due to polypeptide conjugation are not limited to small molecule ligands that bind proteins in the mM to μM range but may be used also for nM ligands to provide recognition elements with dissociation constants in the pM range. Evaluations of two HCA IX constructs that do not carry the proteoglycan (PG) domain did not show significant affinity differences between AZM and the polypeptide conjugate, providing evidence that the improved binding of 4-C10L17-AZM to HCA IX emanated from interactions between the polypeptide segment and the PG domain found only in one carbonic anhydrase, HCA IX. |
| Databáze: | OpenAIRE |
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