Effects of Zn(II) Binding and Apoprotein Structural Stability on the Conformation Change of Designed Antennafinger Proteins

Yuichiro Hori, Yukio Sugiura

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Ligand-induced conformation change is a general strategy for controlling protein function. In this work, we demonstrate the relationships between ligand binding and conformational stability using a previously designed protein, Ant-F, which undergoes a conformation change upon Zn(II) binding. To investigate the effect of stabilization of the apo structure on the conformation change, we also created a novel protein, Ant-F-H1, into which mutations are introduced to increase its stability over that of Ant-F. The chemical denaturation experiments clarified that apo-Ant-F-H1 is more stable than apo-Ant-F (ΔΔG = -1.28 kcal/mol) and that the stability of holo-Ant-F-H1 is almost the same as that of holo-Ant-F. The Zn(II) binding assay shows that the affinity of Zn(II) for Ant-F-H1 is weaker than that for Ant-F (ΔΔG = 1.40 kcal/mol). A large part of the increased value of free energy in stability corresponds to the decreased value of free energy in Zn(II) binding, indicating that the stability of the apo structure directly affects the conformation change. The denaturation experiments also reveal that Zn(II) destabilizes the conformation of both proteins. From the thermodynamic linkage, Zn(II) is thought to bind to the unfolded state with high affinity. These results suggest that the binding of Zn(II) to the unfolded state is an important factor in the conformational change as well as the stability of the apo and holo structures.

Original languageEnglish
Pages (from-to)3068-3074
Number of pages7
JournalBiochemistry
Volume43
Issue number11
DOIs
Publication statusPublished - Mar 23 2004
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry

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