title: Evidence that a metabolic microcompartment contains and recycles private cofactor pools.
abstract: Microcompartments are loose protein cages that encapsulate enzymes for particular bacterial metabolic pathways. These structures are thought to retain and perhaps concentrate pools of small, uncharged intermediates that would otherwise diffuse from the cell. In Salmonella enterica, a microcompartment encloses enzymes for ethanolamine catabolism. The cage has been thought to retain the volatile intermediate acetaldehyde but allow diffusion of the much larger cofactors NAD and coenzyme A (CoA). Genetic tests support an alternative idea that the microcompartment contains and recycles private pools of the large cofactors NAD and CoA. Two central enzymes convert ethanolamine to acetaldehyde (EutBC) and then to acetyl-CoA (EutE). Two seemingly peripheral redundant enzymes encoded by the eut operon proved to be essential for ethanolamine utilization, when subjected to sufficiently stringent tests. These are EutD (acetyl-CoA to acetyl phosphate) and EutG (acetaldehyde to ethanol). Obligatory recycling of cofactors couples the three reactions and drives acetaldehyde consumption. Loss and toxic effects of acetaldehyde are minimized by accelerating its consumption. In a eutD mutant, acetyl-CoA cannot escape the compartment but is released by mutations that disrupt the structure. The model predicts that EutBC (ethanolamine-ammonia lyase) lies outside the compartment, using external coenzyme B12 and injecting its product, acetaldehyde, into the lumen, where it is degraded by the EutE, EutD, and EutG enzymes using private pools of CoA and NAD. The compartment appears to allow free diffusion of the intermediates ethanol and acetyl-PO4 but (to our great surprise) restricts diffusion of acetaldehyde.
authors.affiliation: Department of Microbiology, University of California, Davis, Davis, California, USA.
authors.forename: Douglas L
authors.initials: DL
authors.lastname: Huseby
authors.forename: John R
authors.initials: JR
authors.lastname: Roth
mesh_terms: D000079:Acetaldehyde / Q000378:metabolism; D000105:Acetyl Coenzyme A / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D003065:Coenzyme A / Q000378:metabolism*; D003067:Coenzymes / Q000378:metabolism*; D019856:Ethanolamine / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D008954:Models, Biological; D009243:NAD / Q000378:metabolism*; D019779:Salmonella enterica / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D003067:Coenzymes; D009243:NAD; D019856:Ethanolamine; D000105:Acetyl Coenzyme A; D000079:Acetaldehyde; D003065:Coenzyme A