6H64

Crystal structure of the CRD-SAT

Summary for 6H64

• Entry DOI: 10.2210/pdb6h64/pdb
• Related PRD ID: PRD_900008
• Descriptor: Galectin-3, beta-D-galactopyranose-(1-4)-alpha-D-glucopyranose, SULFATE ION, ... (4 entities in total)
• Functional Keywords: carbohydrate recognition domain, sugar binding protein
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 6
• Total formula weight: 115936.90

Authors

Charron, C.,Kriznik, A.,Yelehe-Okouma, M.,Jouzeau, J.-Y.,Reboul, P. (deposition date: 2018-07-26, release date: 2019-08-14, Last modification date: 2024-01-17)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.8 Å)