8A9Q

Computational design of stable mammalian serum albumins for bacterial expression

8A9Q の概要

• エントリーDOI: 10.2210/pdb8a9q/pdb
• 分子名称: Albumin, MYRISTIC ACID, R-WARFARIN, ... (5 entities in total)
• 機能のキーワード: complex with warfarin, transport protein
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 135874.40

構造登録者

Khersonsky, O.,Dym, O.,Fleishman, J.S. (登録日: 2022-06-29, 公開日: 2023-05-10, 最終更新日: 2024-11-20)

主引用文献

Khersonsky, O.,Goldsmith, M.,Zaretsky, I.,Hamer-Rogotner, S.,Dym, O.,Unger, T.,Yona, M.,Fridmann-Sirkis, Y.,Fleishman, S.J.
Stable Mammalian Serum Albumins Designed for Bacterial Expression.
J.Mol.Biol., 435:168191-168191, 2023

PubMed Abstract: Albumin is the most abundant protein in the blood serum of mammals and has essential carrier and physiological roles. Albumins are also used in a wide variety of molecular and cellular experiments and in the cultivated meat industry. Despite their importance, however, albumins are challenging for heterologous expression in microbial hosts, likely due to 17 conserved intramolecular disulfide bonds. Therefore, albumins used in research and biotechnological applications either derive from animal serum, despite severe ethical and reproducibility concerns, or from recombinant expression in yeast or rice. We use the PROSS algorithm to stabilize human and bovine serum albumins, finding that all are highly expressed in E. coli. Design accuracy is verified by crystallographic analysis of a human albumin variant with 16 mutations. This albumin variant exhibits ligand binding properties similar to those of the wild type. Remarkably, a design with 73 mutations relative to human albumin exhibits over 40 °C improved stability and is stable beyond the boiling point of water. Our results suggest that proteins with many disulfide bridges have the potential to exhibit extreme stability when subjected to design. The designed albumins may be used to make economical, reproducible, and animal-free reagents for molecular and cell biology. They also open the way to high-throughput screening to study and enhance albumin carrier properties.

PubMed: 37385581
DOI: 10.1016/j.jmb.2023.168191

実験手法

X-RAY DIFFRACTION (2 Å)