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Improved molecular recognition of Carbonic Anhydrase IX by polypeptide conjugation to acetazolamide

Yang, Jie; Koruza, Katarina LU ; Fisher, Zoë LU ; Knecht, Wolfgang LU and Baltzer, Lars (2017) In Bioorganic and Medicinal Chemistry
Abstract

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 38. nM and that of the AZM conjugated polypeptide (4-C10L17-AZM) to HCA II was found to be 4. nM, 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 3. nM, whereas that of the 4-C10L17-AZM conjugate was 90. pM, a 33-fold affinity enhancement. This dramatic affinity increase due to polypeptide conjugation was achieved for a small molecule ligand with an... (More)

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 38. nM and that of the AZM conjugated polypeptide (4-C10L17-AZM) to HCA II was found to be 4. nM, 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 3. nM, whereas that of the 4-C10L17-AZM conjugate was 90. pM, 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.

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author
organization
publishing date
type
Contribution to journal
publication status
in press
subject
keywords
Acetazolamide, Human Carbonic Anhydrase, Recognition, Surface plasmon resonance, Tumor
in
Bioorganic and Medicinal Chemistry
publisher
Elsevier
external identifiers
  • scopus:85029715183
  • wos:000413402100064
ISSN
0968-0896
DOI
10.1016/j.bmc.2017.09.017
language
English
LU publication?
yes
id
8a41c343-eb8f-4f82-88a5-4550ede6d2dc
date added to LUP
2017-10-10 07:49:22
date last changed
2018-01-16 13:22:12
@article{8a41c343-eb8f-4f82-88a5-4550ede6d2dc,
  abstract     = {<p>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 38. nM and that of the AZM conjugated polypeptide (4-C10L17-AZM) to HCA II was found to be 4. nM, 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 3. nM, whereas that of the 4-C10L17-AZM conjugate was 90. pM, 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.</p>},
  author       = {Yang, Jie and Koruza, Katarina and Fisher, Zoë and Knecht, Wolfgang and Baltzer, Lars},
  issn         = {0968-0896},
  keyword      = {Acetazolamide,Human Carbonic Anhydrase,Recognition,Surface plasmon resonance,Tumor},
  language     = {eng},
  publisher    = {Elsevier},
  series       = {Bioorganic and Medicinal Chemistry},
  title        = {Improved molecular recognition of Carbonic Anhydrase IX by polypeptide conjugation to acetazolamide},
  url          = {http://dx.doi.org/10.1016/j.bmc.2017.09.017},
  year         = {2017},
}