8QRE

Cholera holotoxin (wildtype)

これはPDB形式変換不可エントリーです。

8QRE の概要

• エントリーDOI: 10.2210/pdb8qre/pdb
• 分子名称: Cholera enterotoxin subunit A, Cholera enterotoxin subunit B, SODIUM ION, ... (9 entities in total)
• 機能のキーワード: cholera toxin, vibrio cholerae, bacterial toxin, vibrio natriegens, holotoxin, toxin
• 由来する生物種: Vibrio cholerae O1; 詳細
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 174336.86

構造登録者

Mojica, N.,Cordara, G.,Krengel, U. (登録日: 2023-10-06, 公開日: 2024-05-15, 最終更新日: 2024-11-20)

主引用文献

Mojica, N.,Kersten, F.,Montserrat-Canals, M.,Huhn Iii, G.R.,Tislevoll, A.M.,Cordara, G.,Teter, K.,Krengel, U.
Using Vibrio natriegens for High-Yield Production of Challenging Expression Targets and for Protein Perdeuteration.
Biochemistry, 63:587-598, 2024

PubMed Abstract: Production of soluble proteins is essential for structure/function studies; however, this usually requires milligram amounts of protein, which can be difficult to obtain with traditional expression systems. Recently, the Gram-negative bacterium emerged as a novel and alternative host platform for production of proteins in high yields. Here, we used a commercial strain derived from (Vmax X2) to produce soluble bacterial and fungal proteins in milligram scale, which we struggled to achieve in . These proteins include the cholera toxin (CT) and -acetyl glucosamine-binding protein A (GbpA) from , the heat-labile enterotoxin (LT) from and the fungal nematotoxin CCTX2 from . CT, GbpA, and LT are secreted by the Type II secretion system in their natural hosts. When these three proteins were produced in Vmax, they were also secreted and could be recovered from the growth media. This simplified the downstream purification procedure and resulted in considerably higher protein yields compared to production in (6- to 26-fold increase). We also tested Vmax for protein perdeuteration using deuterated minimal media with deuterium oxide as solvent and achieved a 3-fold increase in yield compared to the equivalent protocol in . This is good news, since isotopic labeling is expensive and often ineffective but represents a necessary prerequisite for some structural biology techniques. Thus, Vmax represents a promising host for production of challenging expression targets and for protein perdeuteration in amounts suitable for structural biology studies.

PubMed: 38359344
DOI: 10.1021/acs.biochem.3c00612

実験手法

X-RAY DIFFRACTION (2.3 Å)