6E7M

Crystal structure of the holo retinal-bound domain-swapped dimer Q108K:T51D:A28C mutant of human Cellular Retinol Binding Protein II

6E7M の概要

• エントリーDOI: 10.2210/pdb6e7m/pdb
• 分子名称: Retinol-binding protein 2, ACETATE ION, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: retinol, ilbp, protein switch, lipid binding protein
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 64005.65

構造登録者

Ghanbarpour, A.,Geiger, J. (登録日: 2018-07-26, 公開日: 2019-10-16, 最終更新日: 2024-11-13)

主引用文献

Ghanbarpour, A.,Pinger, C.,Esmatpour Salmani, R.,Assar, Z.,Santos, E.M.,Nosrati, M.,Pawlowski, K.,Spence, D.,Vasileiou, C.,Jin, X.,Borhan, B.,Geiger, J.H.
Engineering the hCRBPII Domain-Swapped Dimer into a New Class of Protein Switches.
J.Am.Chem.Soc., 141:17125-17132, 2019

PubMed Abstract: Protein conformational switches or allosteric proteins play a key role in the regulation of many essential biological pathways. Nonetheless, the implementation of protein conformational switches in protein design applications has proven challenging, with only a few known examples that are not derivatives of naturally occurring allosteric systems. We have discovered that the domain-swapped (DS) dimer of hCRBPII undergoes a large and robust conformational change upon retinal binding, making it a potentially powerful template for the design of protein conformational switches. Atomic resolution structures of the apo- and holo-forms illuminate a simple, mechanical movement involving sterically driven torsion angle flipping of two residues that drive the motion. We further demonstrate that the conformational "readout" can be altered by addition of cross-domain disulfide bonds, also visualized at atomic resolution. Finally, as a proof of principle, we have created an allosteric metal binding site in the DS dimer, where ligand binding results in a reversible 5-fold loss of metal binding affinity. The high resolution structure of the metal-bound variant illustrates a well-formed metal binding site at the interface of the two domains of the DS dimer and confirms the design strategy for allosteric regulation.

PubMed: 31557439
DOI: 10.1021/jacs.9b04664

実験手法

X-RAY DIFFRACTION (2.7 Å)