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Andersson, Theresa
Publications (2 of 2) Show all publications
Andersson, T., Lundqvist, M., Dolphin, G. T., Enander, K., Jonsson, B.-H., Nilsson, J. W. & Baltzer, L. (2005). The binding of human Carbonic Anhydrase II by functionalized folded polypeptide receptors. Chemistry and Biology, 12(11), 1245-1252
Open this publication in new window or tab >>The binding of human Carbonic Anhydrase II by functionalized folded polypeptide receptors
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2005 (English)In: Chemistry and Biology, ISSN 1074-5521, E-ISSN 1879-1301, Vol. 12, no 11, p. 1245-1252Article in journal (Refereed) Published
Abstract [en]

Several receptors for human carbonic anhydrase II (HCAII) have been prepared by covalently attaching benzenesulfonamide carboxylates via aliphatic aminocarboxylic acid spacers of variable length to the side chain of a lysine residue in a designed 42 residue helix-loop-helix motif. The sulfonamide group binds to the active site zinc ion of human carbonic anhydrase II located in a 15 Å deep cleft. The dissociation constants of the receptor-HCAII complexes were found to be in the range from low micromolar to better than 20 nM, with the lowest affinities found for spacers with less than five methylene groups and the highest affinity found for the spacer with seven methylene groups. The results suggest that the binding is a cooperative event in which both the sulfonamide residue and the helix-loop-helix motif contribute to the overall affinity.

National Category
Natural Sciences
urn:nbn:se:liu:diva-13359 (URN)10.1016/j.chembiol.2005.08.018 (DOI)
Available from: 2005-09-21 Created: 2005-09-21 Last updated: 2017-12-13
Andersson, T. (2004). Molecular recognition of proteins by functionalized folded polypeptide receptors. (Licentiate dissertation). Linköping: Linköpings universitet
Open this publication in new window or tab >>Molecular recognition of proteins by functionalized folded polypeptide receptors
2004 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis describes the design, synthesis and characterization of synthetic receptor molecules for the recognition and binding of proteins with applications in bioseparation and biosensing. A 42-residue polypeptide, designed to fold into a helix-loop-helix motif and dimerize in solution to form a four-helix bundle, was used as the scaffold. In the first part of the thesis it was functionalized by the incorporation of three substituents at the side chains of lysine residues. A library of 343 receptors was created and screened for affinity towards the human IgG fab fragment using SPR technology. The scaffold was reacted in a stepwise and combinatorial procedure with seven active esters in a pH controlled site-selective acylation reaction to form amides at the side chains of three lysine residues. Four receptor candidates were found to have 0.1 mM affinities and were selected for further investigation.

Both the unfunctionalized scaffold and the four selected receptors were found to bind well also to HCA II and the molecular interactions with this target protein were studied in detail. NMR studies of their interactions with 15N-labeled HCA II revealed that the peptides bound to a hydrophobic patch near the active site cleft, and SPR studies of modified receptor polypeptides led to the conclusion that mainly hydrophobic interactions were involved in binding.

In the second part of the thesis two scaffolds were functionalized with a benzenesulfonamide ligand linked to the scaffold by a series of aliphatic spacers of varying length. Benzenesulfonamide is a known inhibitor of HCA II with a dissociation constant of 1.5 µM and it was found that the overall affinity of the functionalized peptide was enhanced by increasing the length of the ligand spacer due to cooperativity between the scaffold and the ligand in the binding to HCA II. The receptor with a seven methylene group spacer bound HCA II with a dissociation constant of 4 n M. It was also shown that the sequence of the scaffold polypeptide strongly affected the overall affinity of the peptide conjugate for the target protein.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2004. p. 43
Linköping Studies in Science and Technology. Thesis, ISSN 0280-7971 ; 1078
National Category
Cell and Molecular Biology
urn:nbn:se:liu:diva-153049 (URN)LiU-TEK-LIC-2004:07 (Local ID)9173739170 (ISBN)LiU-TEK-LIC-2004:07 (Archive number)LiU-TEK-LIC-2004:07 (OAI)
Available from: 2019-01-14 Created: 2019-01-14 Last updated: 2019-05-10Bibliographically approved

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