Application | Comment | Organism |
---|---|---|
molecular biology | His-tag-induced crystallization of OcDH is found to be dependent on the length of the His tag | Pecten maximus |
Cloned (Comment) | Organism |
---|---|
OcDH is cloned and expressed tagless (OcDH-tagless), as a His5-tagged OcDH (OcDH-His5) and as a His6-tagged OcDH variant (OcDHLEHis6) in Escherichia coli. The His tag is always placed at the C-terminus | Pecten maximus |
Crystallization (Comment) | Organism |
---|---|
His-tag-induced crystallization of OcDH is found to be dependent on the length of the His tag. OcDH-tagless and His6-tagged OcDH do not yield crystals. The purified His5-tagged OcDH yields small crystals. Together with the coenzyme NADH the crystals obtained are single and diffract to 2.1 A resolution. Two distinct domains can be found in OcDH: an NADH-dependent glycerol-3-phosphate dehydrogenase-like domain (domain I) and an octopine dehydrogenase specific domain (domain II; Interpro database). Each domain comprises approximately half of the protein, with domain I containing the classic Rossman fold of dinucleotide-binding proteins | Pecten maximus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pecten maximus | Q9BHM6 | - |
- |
Purification (Comment) | Organism |
---|---|
His5-tagged OcDH and His6-tagged OcDH are purified by Ni-NTA chromatography, yielding almost 20 mg homogenous enzyme per litre of cell culture (purity greater than 98%). The purification of OcDH-tagless requires several chromatographic steps and yields 3-5 mg per litre of cell culture, with a purity of more than 98% | Pecten maximus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
in terms of activity, the three constructs (tagless OcDH, His5-tagged OcDH and His6-tagged OcDH) are indistinguishable | Pecten maximus |
Synonyms | Comment | Organism |
---|---|---|
OcDH | - |
Pecten maximus |
Octopine dehydrogenase | - |
Pecten maximus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Pecten maximus |