4UDM

Crystal structure of Im3 in complex with Y52A mutant of E3RNase

4UDM の概要

• エントリーDOI: 10.2210/pdb4udm/pdb
• 分子名称: COLICIN-E3 IMMUNITY PROTEIN, COLICIN-E3, CALCIUM ION, ... (5 entities in total)
• 機能のキーワード: translation, antibiotic, antimicrobial, bacteriocin, bacteriocin immunity, endonuclease, hydrolase, inhibition, nuclease, protein-protein interactions, ribonuclease, ribosome inactivation, toxin
• 由来する生物種: ESCHERICHIA COLI BL21(DE3); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 20786.47

構造登録者

Sharma, A.,Kleanthous, C. (登録日: 2014-12-10, 公開日: 2016-01-13, 最終更新日: 2023-12-20)

主引用文献

Papadakos, G.,Sharma, A.,Lancaster, L.E.,Bowen, R.,Kaminska, R.,Leech, A.P.,Walker, D.,Redfield, C.,Kleanthous, C.
Consequences of Inducing Intrinsic Disorder in a High-Affinity Protein-Protein Interaction.
J.Am.Chem.Soc., 137:5252-, 2015

PubMed Abstract: The kinetic and thermodynamic consequences of intrinsic disorder in protein-protein recognition are controversial. We address this by inducing one partner of the high-affinity colicin E3 rRNase domain-Im3 complex (K(d) ≈ 10(-12) M) to become an intrinsically disordered protein (IDP). Through a variety of biophysical measurements, we show that a single alanine mutation at Tyr507 within the hydrophobic core of the isolated colicin E3 rRNase domain causes the enzyme to become an IDP (E3 rRNase(IDP)). E3 rRNase(IDP) binds stoichiometrically to Im3 and forms a structure that is essentially identical to the wild-type complex. However, binding of E3 rRNase(IDP) to Im3 is 4 orders of magnitude weaker than that of the folded rRNase, with thermodynamic parameters reflecting the disorder-to-order transition on forming the complex. Critically, pre-steady-state kinetic analysis of the E3 rRNase(IDP)-Im3 complex demonstrates that the decrease in affinity is mostly accounted for by a drop in the electrostatically steered association rate. Our study shows that, notwithstanding the advantages intrinsic disorder brings to biological systems, this can come at severe kinetic and thermodynamic cost.

PubMed: 25856265
DOI: 10.1021/JA512607R

実験手法

X-RAY DIFFRACTION (2.96 Å)