5m42
Structure of Thermus thermophilus L-proline dehydrogenase lacking alpha helices A, B and C
Structural highlights
FunctionPRODH_THET2 Converts proline to delta-1-pyrroline-5-carboxylate (PubMed:17344208, PubMed:18426222). Has significant activity against O(2) producing superoxide during proline oxidation catalytic cycle (PubMed:17344208).[1] [2] Publication Abstract from PubMedFlavoenzymes are versatile biocatalysts containing either FAD or FMN as cofactor. FAD often binds to a Rossmann fold, while FMN prefers a TIM-barrel or flavodoxin-like fold. Proline dehydrogenase is denoted as an exception: it possesses a TIM barrel-like fold while binding FAD. Using a riboflavin auxotrophic Escherichia coli strain and maltose-binding protein as solubility tag, we produced the apoprotein of Thermus thermophilus ProDH (MBP-TtProDH). Remarkably, reconstitution with FAD or FMN revealed that MBP-TtProDH has no preference for either of the two prosthetic groups. Kinetic parameters of both holo forms are similar, as are the dissociation constants for FAD and FMN release. Furthermore, we show that the holo form of MBP-TtProDH, as produced in E. coli TOP10 cells, contains about three times more FMN than FAD. In line with this flavin content, the crystal structure of TtProDH variant DeltaABC, which lacks helices alphaA, alphaB and alphaC, shows no electron density for an AMP moiety of the cofactor. To the best of our knowledge, this is the first example of a flavoenzyme that does not discriminate between FAD and FMN as cofactor. Therefore, classification of TtProDH as an FAD-binding enzyme should be reconsidered. Proline dehydrogenase from Thermus thermophilus does not discriminate between FAD and FMN as cofactor.,Huijbers MM, Martinez-Julvez M, Westphal AH, Delgado-Arciniega E, Medina M, van Berkel WJ Sci Rep. 2017 Mar 3;7:43880. doi: 10.1038/srep43880. PMID:28256579[3] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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