Structural insights into steroid hormone binding

Jarkko Valjakka, Kristiina Takkinen, Tuija Teerinen, Hans Söderlund, Juha Rouvinen

Research output: Contribution to journalArticleScientificpeer-review

18 Citations (Scopus)

Abstract

The monoclonal anti-testosterone antibody (3-C4F5) has a relatively high affinity (3 × 108 M −1) with an overall good specificity profile. However, the earlier characterized binding properties have shown that both the affinity and specificity of this antibody must be improved if it is intended for use in clinical immunoassays. In this paper, the crystal structures of the recombinant anti-testosterone (3-C4F5) Fab fragment have been determined in the testosterone-bound and free form at resolutions of 2.60 and 2.72 Å, respectively. The high affinity binding of the (3-C4F5) Fab is mainly determined by shape complementarity between the protein and testosterone. Only one direct hydrogen bond is formed between the hydroxyl group of the testosterone D-ring and the main-chain oxygen of Gly100JH. The testosterone is deeply bound in a hydrophobic pocket, and the close shape complementarity is mainly formed by the third complementarity-determining regions (CDR) of the heavy and light chain. Comparison of the bound structure with the free structure indicates conformational changes in the protein upon testosterone binding. The conformational changes of the side chains of two residues Glu95H and Tyr99H in the CDR-H3 are particularly essential for the binding. Interesting similarities in the binding of different steroids were also observed upon comparison of the available structures of anti-steroid antibodies.
Original languageEnglish
Pages (from-to)4183-4190
JournalJournal of Biological Chemistry
Volume277
Issue number6
Publication statusPublished - 2002
MoE publication typeA1 Journal article-refereed

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Steroid hormones
Testosterone
Steroids
Hormones
Complementarity Determining Regions
Antibodies
Anti-Idiotypic Antibodies
Immunoglobulin Fab Fragments
Antibody Affinity
Antibody Specificity
Immunoassay
Hydroxyl Radical
Hydrogen
Hydrogen bonds
Proteins
Crystal structure
Monoclonal Antibodies
Oxygen
Light

Cite this

Valjakka, J., Takkinen, K., Teerinen, T., Söderlund, H., & Rouvinen, J. (2002). Structural insights into steroid hormone binding. Journal of Biological Chemistry, 277(6), 4183-4190.
Valjakka, Jarkko ; Takkinen, Kristiina ; Teerinen, Tuija ; Söderlund, Hans ; Rouvinen, Juha. / Structural insights into steroid hormone binding. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 6. pp. 4183-4190.
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Valjakka, J, Takkinen, K, Teerinen, T, Söderlund, H & Rouvinen, J 2002, 'Structural insights into steroid hormone binding', Journal of Biological Chemistry, vol. 277, no. 6, pp. 4183-4190.

Structural insights into steroid hormone binding. / Valjakka, Jarkko; Takkinen, Kristiina; Teerinen, Tuija; Söderlund, Hans; Rouvinen, Juha.

In: Journal of Biological Chemistry, Vol. 277, No. 6, 2002, p. 4183-4190.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Structural insights into steroid hormone binding

AU - Valjakka, Jarkko

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N2 - The monoclonal anti-testosterone antibody (3-C4F5) has a relatively high affinity (3 × 108 M −1) with an overall good specificity profile. However, the earlier characterized binding properties have shown that both the affinity and specificity of this antibody must be improved if it is intended for use in clinical immunoassays. In this paper, the crystal structures of the recombinant anti-testosterone (3-C4F5) Fab fragment have been determined in the testosterone-bound and free form at resolutions of 2.60 and 2.72 Å, respectively. The high affinity binding of the (3-C4F5) Fab is mainly determined by shape complementarity between the protein and testosterone. Only one direct hydrogen bond is formed between the hydroxyl group of the testosterone D-ring and the main-chain oxygen of Gly100JH. The testosterone is deeply bound in a hydrophobic pocket, and the close shape complementarity is mainly formed by the third complementarity-determining regions (CDR) of the heavy and light chain. Comparison of the bound structure with the free structure indicates conformational changes in the protein upon testosterone binding. The conformational changes of the side chains of two residues Glu95H and Tyr99H in the CDR-H3 are particularly essential for the binding. Interesting similarities in the binding of different steroids were also observed upon comparison of the available structures of anti-steroid antibodies.

AB - The monoclonal anti-testosterone antibody (3-C4F5) has a relatively high affinity (3 × 108 M −1) with an overall good specificity profile. However, the earlier characterized binding properties have shown that both the affinity and specificity of this antibody must be improved if it is intended for use in clinical immunoassays. In this paper, the crystal structures of the recombinant anti-testosterone (3-C4F5) Fab fragment have been determined in the testosterone-bound and free form at resolutions of 2.60 and 2.72 Å, respectively. The high affinity binding of the (3-C4F5) Fab is mainly determined by shape complementarity between the protein and testosterone. Only one direct hydrogen bond is formed between the hydroxyl group of the testosterone D-ring and the main-chain oxygen of Gly100JH. The testosterone is deeply bound in a hydrophobic pocket, and the close shape complementarity is mainly formed by the third complementarity-determining regions (CDR) of the heavy and light chain. Comparison of the bound structure with the free structure indicates conformational changes in the protein upon testosterone binding. The conformational changes of the side chains of two residues Glu95H and Tyr99H in the CDR-H3 are particularly essential for the binding. Interesting similarities in the binding of different steroids were also observed upon comparison of the available structures of anti-steroid antibodies.

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