6H64

Crystal structure of the CRD-SAT

6H64 の概要

• エントリーDOI: 10.2210/pdb6h64/pdb
• 関連するBIRD辞書のPRD_ID: PRD_900008
• 分子名称: Galectin-3, beta-D-galactopyranose-(1-4)-alpha-D-glucopyranose, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: carbohydrate recognition domain, sugar binding protein
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 115936.90

構造登録者

Charron, C.,Kriznik, A.,Yelehe-Okouma, M.,Jouzeau, J.-Y.,Reboul, P. (登録日: 2018-07-26, 公開日: 2019-08-14, 最終更新日: 2024-01-17)

主引用文献

Kriznik, A.,Yelehe-Okouma, M.,Lec, J.C.,Groshenry, G.,Le Cordier, H.,Charron, C.,Quinternet, M.,Mazon, H.,Talfournier, F.,Boschi-Muller, S.,Jouzeau, J.Y.,Reboul, P.
CRD SAT Generated by pCARGHO: A New Efficient Lectin-Based Affinity Tag Method for Safe, Simple, and Low-Cost Protein Purification.
Biotechnol J, 14:e1800214-e1800214, 2019

PubMed Abstract: Purification of recombinant proteins remains a bottleneck for downstream processing. The authors engineered a new galectin 3 truncated form (CRD ), functionally and structurally characterized, with preserved solubility and lectinic activity. Taking advantage of these properties, the authors designed an expression vector (pCARGHO), suitable for CRD -tagged protein expression in prokaryotes. CRD binds to lactose-Sepharose with a high specificity and facilitates solubilization of fusion proteins. This tag is structurally stable and can be easily removed from fusion proteins using TEV protease. Furthermore, due to their basic isoelectric point (pI), CRD , and TEV are efficiently eliminated using cationic exchange chromatography. When pI of the protein of interest (POI) and CRD are close, other chromatographic methods are successfully tested. Using CRD tag, the authors purified several proteins from prokaryote and eukaryote origin and demonstrated as examples, the preservation of both Escherichia coli Thioredoxin 1 and human CDC25B activities. Overall, yields of proteins obtained after tag removal are about 5-50 mg per litre of bacterial culture. Our purification method displays various advantages described herein that may greatly interest academic laboratories, biotechnology, and pharmaceutical companies.

PubMed: 30298550
DOI: 10.1002/biot.201800214

実験手法

X-RAY DIFFRACTION (1.8 Å)