Coenzyme- and His-tag-induced crystallization of octopine dehydrogenase

Autor: Sander H. J. Smits, Lutz Schmitt, Manfred K. Grieshaber, Andre Mueller
Rok vydání: 2008
Předmět:
Stereochemistry
chemistry [Recombinant Proteins]
biosynthesis [Recombinant Proteins]
Biophysics
genetics [Amino Acid Oxidoreductases]
Coenzymes
Dehydrogenase
Biology
Crystallography
X-Ray
Biochemistry
Cofactor
law.invention
chemistry.chemical_compound
Protein structure
Structural Biology
law
ddc:570
chemistry [Histidine]
Protein purification
Genetics
Animals
Histidine
Crystallization
Cloning
Molecular
isolation & purification [Amino Acid Oxidoreductases]
chemistry.chemical_classification
Octopine
chemistry [Coenzymes]
Condensed Matter Physics
Recombinant Proteins
Protein Structure
Tertiary
D-octopine dehydrogenase
Crystallography
Pectinidae
Enzyme
chemistry
Crystallization Communications
biology.protein
chemistry [Amino Acid Oxidoreductases]
Amino Acid Oxidoreductases
Protein crystallization
Zdroj: Acta crystallographica / D 64, 836-839 (2008). doi:10.1107/S1744309108025487
ISSN: 1744-3091
DOI: 10.1107/S1744309108025487
Popis: Over the last decade, protein purification has become more efficient and standardized through the introduction of affinity tags. The choice and position of the tag, however, can directly influence the process of protein crystallization. Octopine dehydrogenase (OcDH) without a His tag and tagged protein constructs such as OcDH-His(5) and OcDH-LEHis(6) have been investigated for their crystallizability. Only OcDH-His(5) yielded crystals; however, they were multiple. To improve crystal quality, the cofactor NADH was added, resulting in single crystals that were suitable for structure determination. As shown by the structure, the His(5) tag protrudes into the cleft between the NADH and L-arginine-binding domains and is mainly fixed in place by water molecules. The protein is thereby stabilized to such an extent that the formation of crystal contacts can proceed. Together with NADH, the His(5) tag obviously locks the enzyme into a specific conformation which induces crystal growth.
Databáze: OpenAIRE
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