8DEN

Heme-Free Cytochrome Variant ApoCyt

8DEN の概要

• エントリーDOI: 10.2210/pdb8den/pdb
• 分子名称: Soluble cytochrome b562 (2 entities in total)
• 機能のキーワード: apo cytochrome b562, engineered protein, de novo protein
• 由来する生物種: Escherichia coli BL21(DE3)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 47577.18

構造登録者

Hoffnagle, A.H.,Eng, V.H.,Tezcan, F.A. (登録日: 2022-06-20, 公開日: 2022-07-06, 最終更新日: 2023-10-18)

主引用文献

Hoffnagle, A.M.,Eng, V.H.,Markel, U.,Tezcan, F.A.
Computationally Guided Redesign of a Heme-free Cytochrome with Native-like Structure and Stability.
Biochemistry, 61:2063-2072, 2022

PubMed Abstract: Metals can play key roles in stabilizing protein structures, but ensuring their proper incorporation is a challenge when a metalloprotein is overexpressed in a non-native cellular environment. Here, we have used computational protein design tools to redesign cytochrome (cyt ), which relies on the binding of its heme cofactor to achieve its proper fold, into a stable, heme-free protein. The resulting protein, ApoCyt, features only four mutations and no metal-ligand or covalent bonds, yet displays improved stability over cyt . Mutagenesis studies and X-ray crystal structures reveal that the increase in stability is due to the computationally prescribed mutations, which stabilize the protein fold through a combination of hydrophobic packing interactions, hydrogen bonds, and cation-π interactions. Upon installation of the relevant mutations, ApoCyt is capable of assembling into previously reported, cytochrome-based trimeric and tetrameric assemblies, demonstrating that ApoCyt retains the structure and assembly properties of cyt . The successful design of ApoCyt therefore enables further functional diversification of cytochrome-based assemblies and demonstrates that structural metal cofactors can be replaced by a small number of well-designed, non-covalent interactions.

PubMed: 36106943
DOI: 10.1021/acs.biochem.2c00369

実験手法

X-RAY DIFFRACTION (1.69 Å)