4a2c
Crystal structure of galactitol-1-phosphate dehydrogenase from Escherichia coliCrystal structure of galactitol-1-phosphate dehydrogenase from Escherichia coli
Structural highlights
FunctionGATD_ECOLI Converts galactitol 1-phosphate to tagatose 6-phosphate. Publication Abstract from PubMedEndogenous galactitol-1-phosphate 5-dehydrogenase (GPDH) (EC 1.1.1.251) from Escherichia coli spontaneously interacts with Ni(2+)-NTA matrices becoming a potential contaminant for recombinant, target His-tagged proteins. Purified recombinant, untagged GPDH (rGPDH) converted galactitol into tagatose, and d-tagatose-6-phosphate into galactitol-1-phosphate, in a Zn(2+)- and NAD(H)-dependent manner and readily crystallized what has permitted to solve its crystal structure. In contrast, N-terminally His-tagged GPDH was marginally stable and readily aggregated. The structure of rGPDH revealed metal-binding sites characteristic from the medium-chain dehydrogenase/reductase protein superfamily which may explain its ability to interact with immobilized metals. The structure also provides clues on the harmful effects of the N-terminal His-tag. STRUCTURED SUMMARY OF PROTEIN INTERACTIONS: GPDH and GPDHbind by molecular sieving (View interaction) GPDH and GPDHbind by x-ray crystallography(View interaction) GPDH and GPDHbind by cosedimentation in solution (View interaction). The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes.,Esteban-Torres M, Alvarez Y, Acebron I, Rivas Bde L, Munoz R, Kohring GW, Roa AM, Sobrino M, Mancheno JM FEBS Lett. 2012 Sep 21;586(19):3127-33. doi: 10.1016/j.febslet.2012.07.073. Epub , 2012 Aug 8. PMID:22979983[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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