New ligand-binding sites identified in the crystal structures of β-lactoglobulin complexes with desipramine
© Joanna I. Loch et al. 2022..
The homodimeric β-lactoglobulin belongs to the lipocalin family of proteins that transport a wide range of hydrophobic molecules and can be modified by mutagenesis to develop specificity for novel groups of ligands. In this work, new lactoglobulin variants, FAF (I56F/L39A/M107F) and FAW (I56F/L39A/M107W), were produced and their interactions with the tricyclic drug desipramine (DSM) were studied using X-ray crystallography, calorimetry (ITC) and circular dichroism (CD). The ITC and CD data showed micromolar affinity of the mutants for DSM and interactions according to the classical one-site binding model. However, the crystal structures unambiguously showed that the FAF and FAW dimers are capable of binding DSM not only inside the β-barrel as expected, but also at the dimer interface and at the entrance to the binding pocket. The presented high-resolution crystal structures therefore provide important evidence of the existence of alternative ligand-binding sites in the β-lactoglobulin molecule. Analysis of the crystal structures highlighted the importance of shape complementarity for ligand recognition and selectivity. The binding sites identified in the crystal structures of the FAF-DSM and FAW-DSM complexes together with data from the existing literature are used to establish a systematic classification of the ligand-binding sites in the β-lactoglobulin molecule.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2022 |
|---|---|
| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:9 |
|---|---|
| Enthalten in: |
IUCrJ - 9(2022), Pt 3 vom: 01. Mai, Seite 386-398 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Loch, Joanna I [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
β-lactoglobulin |
|---|
| Anmerkungen: |
Date Revised 16.07.2022 published: Electronic-eCollection Citation Status PubMed-not-MEDLINE |
|---|
| doi: |
10.1107/S2052252522004183 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM34074958X |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM34074958X | ||
| 003 | DE-627 | ||
| 005 | 20231226010157.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2022 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1107/S2052252522004183 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1135.xml |
| 035 | |a (DE-627)NLM34074958X | ||
| 035 | |a (NLM)35546795 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Loch, Joanna I |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a New ligand-binding sites identified in the crystal structures of β-lactoglobulin complexes with desipramine |
| 264 | 1 | |c 2022 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Revised 16.07.2022 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status PubMed-not-MEDLINE | ||
| 520 | |a © Joanna I. Loch et al. 2022. | ||
| 520 | |a The homodimeric β-lactoglobulin belongs to the lipocalin family of proteins that transport a wide range of hydrophobic molecules and can be modified by mutagenesis to develop specificity for novel groups of ligands. In this work, new lactoglobulin variants, FAF (I56F/L39A/M107F) and FAW (I56F/L39A/M107W), were produced and their interactions with the tricyclic drug desipramine (DSM) were studied using X-ray crystallography, calorimetry (ITC) and circular dichroism (CD). The ITC and CD data showed micromolar affinity of the mutants for DSM and interactions according to the classical one-site binding model. However, the crystal structures unambiguously showed that the FAF and FAW dimers are capable of binding DSM not only inside the β-barrel as expected, but also at the dimer interface and at the entrance to the binding pocket. The presented high-resolution crystal structures therefore provide important evidence of the existence of alternative ligand-binding sites in the β-lactoglobulin molecule. Analysis of the crystal structures highlighted the importance of shape complementarity for ligand recognition and selectivity. The binding sites identified in the crystal structures of the FAF-DSM and FAW-DSM complexes together with data from the existing literature are used to establish a systematic classification of the ligand-binding sites in the β-lactoglobulin molecule | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a desipramine | |
| 650 | 4 | |a ligand-binding sites | |
| 650 | 4 | |a β-lactoglobulin | |
| 700 | 1 | |a Barciszewski, Jakub |e verfasserin |4 aut | |
| 700 | 1 | |a Śliwiak, Joanna |e verfasserin |4 aut | |
| 700 | 1 | |a Bonarek, Piotr |e verfasserin |4 aut | |
| 700 | 1 | |a Wróbel, Paulina |e verfasserin |4 aut | |
| 700 | 1 | |a Pokrywka, Kinga |e verfasserin |4 aut | |
| 700 | 1 | |a Shabalin, Ivan G |e verfasserin |4 aut | |
| 700 | 1 | |a Minor, Wladek |e verfasserin |4 aut | |
| 700 | 1 | |a Jaskolski, Mariusz |e verfasserin |4 aut | |
| 700 | 1 | |a Lewiński, Krzysztof |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t IUCrJ |d 2014 |g 9(2022), Pt 3 vom: 01. Mai, Seite 386-398 |w (DE-627)NLM240540468 |x 2052-2525 |7 nnns |
| 773 | 1 | 8 | |g volume:9 |g year:2022 |g number:Pt 3 |g day:01 |g month:05 |g pages:386-398 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1107/S2052252522004183 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 9 |j 2022 |e Pt 3 |b 01 |c 05 |h 386-398 | ||